Novel Polynucleotides
Novel polynucleotides derived from microorganisms belonging to coryneform bacteria and fragments thereof, polypeptides encoded by the polynucleotides and fragments thereof, polynucleotide arrays comprising the polynucleotides and fragments thereof, recording media in which the nucleotide sequences of the polynucleotide and fragments thereof have been recorded which are readable in a computer, and use of them.
[0001] The present application claims benefit of Japanese Patent Application Nos. Hei. 11-377484 (filed Dec. 16, 1999), 2000-159162 (filed Apr. 7, 2000) and 2000-280988 (filed Aug. 3, 2000), the entire contents of each of which is incorporated herein by reference.
[0002] The contents of the attached CD-R compact discs are incorporated herein by reference in their entirety. The attached discs contain an identical copy of a file “SEQ2.TXT” which were created on the discs on Dec. 13, 2000, and are each 25,891 KR.
BACKGROUND OF THE INVENTION[0003] 1. Field of the Invention
[0004] The present invention relates to novel polynucleotides derived from microorganisms belonging to coryneform bacteria and fragments thereof, polypeptides encoded by the polynucleotides and fragments thereof, polynucleotide arrays comprising the polynucleotides and fragments thereof, computer readable recording media in which the nucleotide sequences of the polynucleotide and fragments thereof have been recorded, and use of them as well as a method of using the polynucleotide and/or polypeptide sequence information to make comparisons.
[0005] 2. Brief Description of the Background Art
[0006] Coryneform bacteria are used in producing various useful substances, such as amino acids, nucleic acids, vitamins, saccharides (for example, ribulose), organic acids (for example, pyruvic acid), and analogues of the above-described substances (for example, N-acetylamino acids) and are very useful microorganisms industrially. Many mutants thereof are known.
[0007] For example, Corynebacterium glutamicum is a Gram-positive bacterium identified as a glutamic acid-producing bacterium, and many amino acids are produced by mutants thereof. For example, 1,000,000 ton/year of L-glutamic acid which is useful as a seasoning for umami (delicious taste), 250,000 ton/year of L-lysine which is a valuable additive for livestock feeds and the like, and several hundred ton/year or more of other amino acids, such as L-arginine, L-proline, L-glutamine, L-tryptophan, and the like, have been produced in the world (Nikkei Bio Yearbook 99, published by Nikkei BP (1998)).
[0008] The production of amino acids by Corynebacterium glutamicum is mainly carried out by its mutants (metabolic mutants) which have a mutated metabolic pathway and regulatory systems. In general, an organism is provided with various metabolic regulatory systems so as not to produce more amino acids than it needs. In the biosynthesis of L-lysine, for example, a microorganism belonging to the genus Corynebacterium is under such regulation as preventing the excessive production by concerted inhibition by lysine and threonine against the activity of a biosynthesis enzyme common to lysine, threonine and methionine, i.e., an aspartokinase, (J. Biochem., 65:849-859 (1969)). The biosynthesis of arginine is controlled by repressing the expression of its biosynthesis gene by arginine so as not to biosynthesize an excessive amount of arginine (Microbiology, 142:99-108 (1996)) It is considered that these metabolic regulatory mechanisms are deregulated in amino acid-producing mutants. Similarly, the metabolic regulation is deregulated in mutants producing nucleic acids, vitamins, saccharides, organic acids and analogues of the above-described substances so as to improve the productivity of the objective product.
[0009] However, accumulation of basic genetic, biochemical and molecular biological data on coryneform bacteria is insufficient in comparison with Escherichia coli, Bacillus subtilis, and the like. Also, few findings have been obtained on mutated genes in amino acid-producing mutants. Thus, there are various mechanisms, which are still unknown, of regulating the growth and metabolism of these microorganisms.
[0010] A chromosomal physical map of Corynebacterium glutamicum ATCC 13032 is reported and it is known that its genome size is about 3,100 kb (Mol. Gen. Genet., 252:255-265 (1996)). Calculating on the basis of the usual gene density of bacteria, it is presumed that about 3,000 genes are present in this genome of about 3,100 kb. However, only about 100 genes mainly concerning amino acid biosynthesis genes are known in Corynebacterium glutamicum, and the nucleotide sequences of most genes have not been clarified hitherto.
[0011] In recent years, the full nucleotide sequence of the genomes of several microorganisms, such as Escherichia coli, Mycobacterium tuberculosis, yeast, and the like, have been determined (Science, 277:1453-62 (1997); Nature, 393:537-544 (1998); Nature, 387. 5-105 (1997)). Based on the thus determined full nucleotide sequences, assumption of gene regions and prediction of their function by comparison with the nucleotide sequences of known genes have been carried out. Thus, the functions of a great number of genes have been presumed, without genetic, biochemical or molecular biological experiments.
[0012] In recent years, moreover, techniques for monitoring expression levels of a great number of genes simultaneously or detecting mutations, using DNA chips, DNA arrays or the like in which a partial nucleic acid fragment of a gene or a partial nucleic acid fragment in genomic DNA other than a gene is fixed to a solid support, have been developed. The techniques contribute to the analysis of microorganisms, such as yeasts, Mycobacterium tuberculosis, Mycobacterium bovis used in BCG vaccines, and the like (Science, 278:680-686 (1997); Proc. Natl. Acad. Sci. USA, 96:12833-38 (1999); Science, 284:1520-23 (1999)).
SUMMARY OF THE INVENTION[0013] An object of the present invention is to provide a polynucleotide and a polypeptide derived from a microorganism of coryneform bacteria which are industrially useful, sequence information of the polynucleotide and the polypeptide, a method for analyzing the microorganism, an apparatus and a system for use in the analysis, and a method for breeding the microorganism.
[0014] The present invention provides a polynucleotide and an oligonucleotide derived from a microorganism belonging to coryneform bacteria, oligonucleotide arrays to which the polynucleotides and the oligonucleotides are fixed, a polypeptide encoded by the polynucleotide, an antibody which recognizes the polypeptide, polypeptide arrays to which the polypeptides or the antibodies are fixed, a computer readable recording medium in which the nucleotide sequences of the polynucleotide and the oligonucleotide and the amino acid sequence of the polypeptide have been recorded, and a system based on the computer using the recording medium as well as a method of using the polynucleotide and/or polypeptide sequence information to make comparisons.
BRIEF DESCRIPTION OF THE DRAWING[0015] FIG. 1 is a map showing the positions of typical genes on the genome of Corynebacterium glutamicum ATCC 13032.
[0016] FIG. 2 is electrophoresis showing the results of proteome analyses using proteins derived from (A) Corynebacterium glutamicum ATCC 13032, (B) FERM BP-7134, and (C) FERM BP-158.
[0017] FIG. 3 is a flow chart of an example of a system using the computer readable media according to the present invention.
[0018] FIG. 4 is a flow chart of an example of a system using the computer readable media according to the present invention.
DETAILED DESCRIPTION OF THE INVENTION[0019] This application is based on Japanese applications No. Hei. 11-377484 filed on Dec. 16, 1999, No. 2000-159162 filed on Apr. 7, 2000 and No. 2000-280988 filed on Aug. 3, 2000, the entire contents of which are incorporated hereinto by reference.
[0020] From the viewpoint that the determination of the full nucleotide sequence of Corynebacterium glutamicum would make it possible to specify gene regions which had not been previously identified, to determine the function of an unknown gene derived from the microorganism through comparison with nucleotide sequences of known genes and amino acid sequences of known genes, and to obtain a useful mutant based on the presumption of the metabolic regulatory mechanism of a useful product by the microorganism, the inventors conducted intensive studies and, as a result, found that the complete genome sequence of Corynebacterium glutamicum can be determined by applying the whole genome shotgun method.
[0021] Specifically, the present invention relates to the following (1) to (65):
[0022] (1) A method for at least one of the following:
[0023] (A) identifying a mutation point of a gene derived from a mutant of a coryneform bacterium,
[0024] (B) measuring an expression amount of a gene derived from a coryneform bacterium,
[0025] (C) analyzing an expression profile of a gene derived from a coryneform bacterium,
[0026] (D) analyzing expression patterns of genes derived from a coryneform bacterium, or
[0027] (E) identifying a gene homologous to a gene derived from a coryneform bacterium,
[0028] said method comprising:
[0029] (a) producing a polynucleotide array by adhering to a solid support at least two polynucleotides selected from the group consisting of first polynucleotides comprising the nucleotide sequence represented by any one of SEQ ID NOS:1 to 3501, second polynucleotides which hybridize with the first polynucleotides under stringent conditions, and third polynucleotides comprising a sequence of 10 to 200 continuous bases of the first or second polynucleotides,
[0030] (b) incubating the polynucleotide array with at least one of a labeled polynucleotide derived from a coryneform bacterium, a labeled polynucleotide derived from a mutant of the coryneform bacterium or a labeled polynucleotide to be examined, under hybridization conditions,
[0031] (c) detecting any hybridization, and
[0032] (d) analyzing the result of the hybridization.
[0033] As used herein, for example, the at least two polynucleotides can be at least two of the first polynucleotides, at least two of the second polynucleotides, at least two of the third polynucleotides, or at least two of the first, second and third polynucleotides.
[0034] (2) The method according to (1), wherein the coryneform bacterium is a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterinum.
[0035] (3) The method according to (2), wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacteriunm herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
[0036] (4) The method according to (1), wherein the polynucleotide derived from a coryneform bacterium, the polynucleotide derived from a mutant of the coryneform bacterium or the polynucleotide to be examined is a gene relating to the biosynthesis of at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof.
[0037] (5) The method according to (1), wherein the polynucleotide to be examined is derived from Escherichia coli.
[0038] (6) A polynucleotide array, comprising:
[0039] at least two polynucleotides selected from the group consisting of first polynucleotides comprising the nucleotide sequence represented by any one of SEQ ID NOS:1 to 3501, second polynucleotides which hybridize with the first polynucleotides under stringent conditions, and third polynucleotides comprising 10 to 200 continuous bases of the first or second polynucleotides, and
[0040] a solid support adhered thereto.
[0041] As used herein, for example, the at least two polynucleotides can be at least two of the first polynucleotides, at least two of the second polynucleotides, at least two of the third polynucleotides, or at least two of the first, second and third polynucleotides.
[0042] (7) A polynucleotide comprising the nucleotide sequence represented by SEQ ID NO:1 or a polynucleotide having a homology of at least 80% with the polynucleotide.
[0043] (8) A polynucleotide comprising any one of the nucleotide sequences represented by SEQ ID NOS:2 to 3431, or a polynucleotide which hybridizes with the polynucleotide under stringent conditions.
[0044] (9) A polynucleotide encoding a polypeptide having any one of the amino acid sequences represented by SEQ ID NOS:3502 to 6931, or a polynucleotide which hybridizes therewith under stringent conditions.
[0045] (10) A polynucleotide which is present in the 5′ upstream or 3′ downstream of a polynucleotide comprising the nucleotide sequence of any one of SEQ ID NOS:2 to 3431 in a whole polynucleotide comprising the nucleotide sequence represented by SEQ ID NO:1, and has an actionty of regulating an expression of the polynucleotide.
[0046] (11) A polynucleotide comprising 10 to 200 continuous bases in the nucleotide sequence of the polynucleotide of any one of (7) to (10), or a polynucleotide comprising a nucleotide sequence complementary to the polynucleotide comprising 10 to 200 continuous based.
[0047] (12) A recombinant DNA comprising the polynucleotide of any one of (8) to (11).
[0048] (13) A transformant comprising the polynucleotide of any one of (8) to (11) or the recombinant DNA of (12). (14) A method for producing a polypeptide, comprising:
[0049] culturing the transformant of (13) in a medium to produce and accumulate a polypeptide encoded by the polynucleotide of (8) or (9) in the medium, and
[0050] recovering the polypeptide from the medium.
[0051] (15) A method for producing at least one of an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof, comprising:
[0052] culturing the transformant of (13) in a medium to produce and accumulate at least one of an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof in the medium, and
[0053] recovering the at least one of the amino acid, the nucleic acid, the vitamin, the saccharide, the organic acid, and analogues thereof from the medium.
[0054] (16) A polypeptide encoded by a polynucleotide comprising the nucleotide sequence selected from SEQ ID NOS:2 to 3431.
[0055] (17) A polypeptide comprising the amino acid sequence selected from SEQ ID NOS:3502 to 6931.
[0056] (18) The polypeptide according to (16) or (17), wherein at least one amino acid is deleted, replaced, inserted or added, said polypeptides having an activity which is substantially the same as that of the polypeptide without said at least one amino acid deletion, replacement, insertion or addition.
[0057] (19) A polypeptide comprising an amino acid sequence having a homology of at least 60% with the amino acid sequence of the polypeptide of (16) or (17), and having an activity which is substantially the same as that of the polypeptide.
[0058] (20) An antibody which recognizes the polypeptide of any one of (16) to (19).
[0059] (21) A polypeptide array, comprising:
[0060] at least one polypeptide or partial fragment polypeptide selected from the polypeptides of (16) to (19) and partial fragment polypeptides of the polypeptides, and
[0061] a solid support adhered thereto.
[0062] (22) A polypeptide array, comprising:
[0063] at least one antibody which recognizes a polypeptide or partial fragment polypeptide selected from the polypeptides of (16) to (19) and partial fragment polypeptides of the polypeptides, and
[0064] a solid support adhered thereto.
[0065] (23) A system based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
[0066] (i) a user input device that inputs at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501, and target sequence or target structure motif information;
[0067] (ii) a data storage device for at least temporarily storing the input information;
[0068] (iii) a comparator that compares the at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501 with the target sequence or target structure motif information, recorded by the data storage device for screening and analyzing nucleotide sequence information which is coincident with or analogous to the target sequence or target structure motif information; and
[0069] (iv) an output device that shows a screening or analyzing result obtained by the comparator.
[0070] (24) A method based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
[0071] (i) inputting at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501, target sequence information or target structure motif information into a user input device;
[0072] (ii) at least temporarily storing said information;
[0073] (iii) comparing the at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501 with the target sequence or target structure motif information; and
[0074] (iv) screening and analyzing nucleotide sequence information which is coincident with or analogous to the target sequence or target structure motif information.
[0075] (25) A system based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
[0076] (i) a user input device that inputs at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, and target sequence or target structure motif information;
[0077] (ii) a data storage device for at least temporarily storing the input information;
[0078] (iii) a comparator that compares the at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target sequence or target structure motif information, recorded by the data storage device for screening and analyzing amino acid sequence information which is coincident with or analogous to the target sequence or target structure motif information; and
[0079] (iv) an output device that shows a screening or analyzing result obtained by the comparator.
[0080] (26) A method based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
[0081] (i) inputting at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, and target sequence information or target structure motif information into a user input device;
[0082] (ii) at least temporarily storing said information;
[0083] (iii) comparing the at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target sequence or target structure motif information; and
[0084] (iv) screening and analyzing amino acid sequence information which is coincident with or analogous to the target sequence or target structure motif information.
[0085] (27) A system based on a computer for determining a function of a polypeptide encoded by a polynucleotide having a target nucleotide sequence derived from a coryneform bacterium, comprising the following:
[0086] (i) a user input device that inputs at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501, function information of a polypeptide encoded by the nucleotide sequence, and target nucleotide sequence information;
[0087] (ii) a data storage device for at least temporarily storing the input information;
[0088] (iii) a comparator that compares the at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501 with the target nucleotide sequence information, and determining a function of a polypeptide encoded by a polynucleotide having the target nucleotide sequence which is coincident with or analogous to the polynucleotide having at least one nucleotide sequence selected from SEQ ID NOS:2 to 3501; and
[0089] (iv) an output devices that shows a function obtained by the comparator.
[0090] (28) A method based on a computer for determining a function of a polypeptide encoded by a polypeptide encoded by a polynucleotide having a target nucleotide sequence derived from a coryneform bacterium, comprising the following:
[0091] (i) inputting at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501, function information of a polypeptide encoded by the nucleotide sequence, and target nucleotide sequence information;
[0092] (ii) at least temporarily storing said information;
[0093] (iii) comparing the at least one nucleotide sequence information selected from SEQ ID NoS:2 to 3501 with the target nucleotide sequence information; and
[0094] (iv) determining a function of a polypeptide encoded by a polynucleotide having the target nucleotide sequence which is coincident with or analogous to the polynucleotide having at least one nucleotide sequence selected from SEQ ID NOS:2 to 3501.
[0095] (29) A system based or a computer for determining a function of a polypeptide having a target amino acid sequence derived from a coryneform bacterium, comprising the following:
[0096] (i) a user input device that inputs at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, function information based on the amino acid sequence, and target amino acid sequence information;
[0097] (ii) a data storing device for at least temporarily storing the input information;
[0098] (iii) a comparator that compares the at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target amino acid sequence information for determining a function of a polypeptide having the target amino acid sequence which is coincident with or analogous to the polypeptide having at least one amino acid sequence selected from SEQ ID NOS-3502 to 7001; and
[0099] (iv) an output device that shows a function obtained by the comparator.
[0100] (30) A method based on a computer for determining a function of a polypeptide having a target amino acid sequence derived from a coryneform bacterium, comprising the following:
[0101] (i) inputting at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, function information based on the amino acid sequence, and target amino acid sequence information;
[0102] (ii) at least temporarily storing said information;
[0103] (iii) comparing the at least one amino acid sequence information selected from SEQ ID NOS:3502 to/001 with the target amino acid sequence information; and
[0104] (iv) determining a function of a polypeptide having the target amino acid sequence which is coincident with or analogous to the polypeptide having at least one amino acid sequence selected from SEQ ID NOS:3502 to 7001.
[0105] (31) The system according to any one of (23), (25), (27) and (29), wherein a coryneform bacterium is a microorganism of the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterinum
[0106] (32) The method according to any one of (24), (26), (28) and (30), wherein a coryneform bacterium is a microorganism of the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterinum.
[0107] (33) The system according to (31), wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium giutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
[0108] (34) The method according to (32), wherein the microorganism belonging to the genus Corynebacterium is selected from the grow consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
[0109] (35) A recording medium or storage device which is readable by a computer is which at least one nucleotide sequence information or selected from SEQ ID NOS:1 to 3501 or function information based or the nucleotide sequence is recorded, and is usuable in the system of (23) or (27) or the method of (24) or (28)
[0110] (36) A recording medium or storage device which is readable by a computer in which at least one amino acid sequence information selected from SEQ ID NOS:2502 to 7001 or function information based on the amino acid sequence is recorded, and is usable in the system of (25) or (29) or the method of (26) or (30).
[0111] (37) The recording medium or storage device according to (35) or (36), wherein is a computer readable recording medium selected from the group consisting of a floppy disc, a hard disc, a magnetic tape, a random access memory (RAM), a read only memory (ROM), a magneto-optic disc (MO), CD-ROM, CD-R, CD-RW, DVD-ROM, DVD-RAM and DVD-RW.
[0112] (38) A polypepide having a homoserine dehydrogenase activity, comprising an amino acid sequence in which the Val residue at the sequence in the amino acid sequence of homoserine dehydrogenase derived from a coryneform bacterium is replaced with amino acid residue other than a Val residue.
[0113] (39) A polypeptide comprising an amino acid sequence in which the Val residue at the 59th position at the amino acid sequence as represented by SEQ ID NO:6957 is replaced with amino acid residue other than a Val residue.
[0114] (40) The polypeptide according to (38) or (39) wherein the Val residue at the 59th position is replaced with an Val residue.
[0115] (41) A polypeptide having private carboxylase activity, comprising an amino acid sequence in which the Pro residue at the 458th position in the amino acid sequence of private carboxylase derived from a coryneform bacterium is replaced with an amino acid residue other than a Pro residue.
[0116] (42) A polypeptide cruising an amino acid sequence in which the Pro residue at the 458th position in the amino acid sequence represented by SEQ ID NO-4265 is replaced with an amino acad residue other than a Pro residue.
[0117] (43) The polypeptide according to (41) or (42), wherein the Pro residue at the 458th position is replaced with a Ser residue.
[0118] (44) The polypeptide recording to any one of (38) to (43), which is derived from Corynebacterium bacterium.
[0119] (45) A DNA encoding the polypeptide of any one of (38) to (44).
[0120] (46) A recombinant DNA comprising the DNA of (45).
[0121] (47) A transformant comprising the recombinant DNA of (46).
[0122] (48) A transformant comprising in its chromosome the DNA of (45).
[0123] (49) The transformant according to (47) or (48), which is derived from a coryneform bacterium.
[0124] (50) The transformant according to (49), which is derived from Corynebacterium glutamicum.
[0125] (51) A method for producing L-lysine, comprising:
[0126] culturing the transformant of any one of (47) to
[0127] (50) in a medium to produce and accumulate L-lysine in the medium, and
[0128] recovering the L-lysine from the culture.
[0129] (52) A method for breeding a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:1 to 3431, comprising the following:
[0130] (i) comparing a nucleotide sequence of a genome or gene of a production strain derived a coryneform bacterium which has been subjected to mutation breeding 50 as the produce at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogous thereon by a fermentation method with a corresponding nucleotide sequence in SEQ ID NOS:1 to 3431;
[0131] (ii) identifying a mutation point present in the production strain based on a result obtained by the
[0132] (iii) introducing the mutation point into a coryneform bacterium which is free of the mutation point; and
[0133] (iv) examining productivity by the fermentation method of the compound selected in (i) of the coryneform bacterium obtained in (iii)
[0134] (53) The method according to (52), wherein the gene is a gene encoding an enzyme in a biosynthesis pathway or a signal transmission pathway.
[0135] (54) The method according to (52), wherein the mutation point is a mutation point relating to a useful mutation which improves or stabilizes the productivity.
[0136] (55) A method for breading a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:1 to 3431, comprising:
[0137] (i) comparing a nucleotide sequence of a genome or gene of a production strain derived a coryneform bacterium which has been subjected to mutation breeding so as to produce at least one compound selected from an amino acid, a nucleic acid, a vitamin a saccharide, an organic acid, and analogous thereof by a fermentation method, with a corresponding nucleon of sequence in SEQ ID NOS:1 to 3431;
[0138] (ii) identifying a information pressure in the production strain based on the result obtain by (1);
[0139] (iii) deleting a mutation point from a coryneform bacterium having the mutation point; and
[0140] (iv) examining productivity by the fermentation method of the compound selected in (i) of the coryneform bacterium obtained in (iii).
[0141] (56) The method according to (55), wherein the gene is a gene encoding an enzyme in a biosynthesis pathway or a signal transmission pathway.
[0142] (57) The method according to (55), wherein the mutation point is a mutation point which decreases or destabilizes the productivity.
[0143] (58) A method for breeding a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:2 to 3431, comprising the following:
[0144] (i) identifying an isozyme relating to biosynthesis of at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogous thereof, based on the nucleotide sequence information represented by SEQ ID NOS:2 to 3431;
[0145] (ii) classifying the isozyme identified in (i) into an isozyme having the same activity;
[0146] (iii) mutating all genes encoding the isozyme having the same activity simultaneously; and
[0147] (iv) examining productivity by a fermentation method of the compound selected an (i) of the coryneform bacterium which have been transformed with the gene obtained in (iii).
[0148] (59) A method for breeding a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:2 to 3431, comprising the following:
[0149] (i) arranging a function information of an open reading frame (ORF) represented by SEQ ID NOS:2 to 3431;
[0150] (ii) allowing the arranged ORF to correspond to an enzyme on a known biosynthesis or signal transmission pathway;
[0151] (iii) explicating an unknown biosynthesis pathway or signal transmission pathway of a coryneform bacterium in combination with information relating known biosynthesis pathway or signal transmission pathway of a coryneform bacterium;
[0152] (iv) comparing the pathway explicated in (iii) with a biosynthesis pathway of a target useful product; and
[0153] (v) transgenetically varying a coryneform bacterium based on the nucleotide sequence information to either strengthen a pathway which is judged to be important in the biosynthesis of the target useful product in (iv) or weaken a pathway which is judged not to be important in the biosynthesis of the target useful product in (iv).
[0154] (60) A coryneform bacterium, bred by the method of any one of (52) to (59).
[0155] (61) The coryneform bacterium according to (60), which is a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterinum.
[0156] (62) The coryneform bacterium according to (61), wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
[0157] (63) A method for producing at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid and an analogue thereof, comprising:
[0158] culturing a coryneform bacterium of any one of (60) to (62) in a medium to produce and accumulate at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof;
[0159] recovering the compound from the culture
[0160] (64) The method according to (63), wherein the compound is L-lysine.
[0161] (65) A method for identifying a protein relating to useful mutation based on proteome analysis, comprising the following:
[0162] (i) preparing
[0163] a protein derived from a bacterium of a production strain of a coryneform bacterium which has been subjected to mutation breeding by a fermentation process so as to produce at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof, and
[0164] a protein derived from a bacterium of a parent strain of the production strain;
[0165] (ii) separating the proteins prepared in (i) by two dimensional electrophoresis;
[0166] (iii) detecting the separated proteins, and comparing an expression amount of the protein derived from the production strain with that derived from the parent strain;
[0167] (iv) treating the protein showing different expression amounts as a result of -the comparison with a peptidase to extract peptide fragments;
[0168] (v) analyzing amino acid sequences of the peptide fragments obtained in (iv); and
[0169] (vi) comparing the amino acid sequences obtained in (v) with the amino acid sequence represented by 920 ID NOS:3502 to 7001 to identifying the protein having the amino acid sequences.
[0170] As used herein, the term “proteome”, which is a coined word by combining “protein” with “genome”, refers to a method for examining of a gene at the polypeptide level.
[0171] (66) The method according to (65), wherein the coryneform bacterium is a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterinum.
[0172] (67) The method according to (66), wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
[0173] (68) A biologically pure culture of Corynebacterium glutamicum AHP-3 (FERM BP-7382).
[0174] The present invention will be described below in more detail, based on the determination of the full nucleotide sequence of coryneform bacteria.
[0175] 1. Determination of full nucleotide sequence of coryneform bacteria
[0176] The term “coryneform bacteria” as used herein means a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium or the genus Microbacterinum as defined in Bergeys of Determination Bacteriology, 8:599 (1974)
[0177] Examples include Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium glutamicum, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, Brevibacterium saccharolyticum, Brevibacterium immariophilum, Brevibacterium roseum, Brevibacterium thiogenitalis, Microbacterinum ammoniaphilum, and the like.
[0178] Specific examples include Corynebacterium acetoacidophilum ATCC 13870, Corynebacterium acetoglutamicum ATCC 15806, Corynebacterium callunae ATCC 15991, Corynebacterium glutamicum ATCC 13032, Corynebacterium glutamicum ATCC 13060, Corynebacterium glutamicum ATCC 13826 (prior genus and species: Brevibacterium flavum, or Corynebacterium lactofermentum), Corynebacterium glutamicum ATCC 14020 (prior genus and species:Breribacterium divaricatum), Corynebacterium glutamicum ATCC 13869 (prior genus and species: Brevibacterium lactofermentum), Corynebacterium herculis ATCC 12868, Corynebacterium lilium ATCC 15990, Corynebacteriunm melassecola ATCC 17965, Corynebacterium thermoaminogenes FERM 9244, Brevibacterium saccharolyticum ATCC 14066, Brevibacterium immariophilum ATCC 14068, Brevibacterium roseum ATCC 13825, Brevibacterium thiogenitalis ATCC 19240, Microbacterinum ammoniaphilum ATCC 15354, and the like.
[0179] (1) Preparation of genome DNA of coryneform bacteria
[0180] Coryneform bacteria can be cultured by a conventional method.
[0181] Any of a natural medium and a synthetic medium can be used, so long as it is a medium suitable for efficient culturing of the microorganism, and it contains a carbon source, a nitrogen source, an inorganic salt, and the like which can be assimilated by the microorganism.
[0182] In Corynebacterium glutamicum, for example, a BY medium (7 g/l meat extract, 10 g/l peptone, 3 g/l sodium chloride, 5 g/l yeast extract, pH 7.2) containing 1% of glycine and the like can be used. The culturing is carried out at 25 to 35° C. overnight.
[0183] After the completion of the culture, the cells are recovered from the culture by centrifugation. The resulting cells are washed with a washing solution.
[0184] Examples of the washing solution include STE buffer (10.3% sucrose, 25 mol/l Tris hydrochloride, 25 mmol/l ethylenediaminetetraacetic acid (hereinafter referred to as “EDTA”), pH 8.0), and the like.
[0185] Genome DNA can be obtained from the washed cells according to a conventional method for obtaining genome DNA, namely, lysing the cell wall of the cells using a lysozyme and a surfactant (SDS, etc.), eliminating proteins and the like using a phenol solution and a phenol/chloroform solution, and then precipitating the genome DNA with ethanol or the likes Specifically, the following method can be illustrated.
[0186] The washed cells are suspended in a washing solution containing 5 to 20 mg/l lysozyme. After shaking, 5 to 20% SDS is added to lyse the cells. In usual, shaking is gently performed at 25 to 40° C. for 30 minutes to 2 hours. After shaking, the suspension is maintained at 60 to 70° C. for 5 to 15 minutes for the lysis.
[0187] After the lysis, the suspension is cooled to ordinary temperature, and 5 to 20 ml of Tris-neutralized phenol is added thereto, followed by gently shaking at room temperature for 15 to 45 minutes.
[0188] After shaking, centrifugation (15,000×g, 20 minutes, 20° C.) is carried out to fractionate the aqueous layer.
[0189] After performing extraction with phenol/chloroform and extraction with chloroform (twice) in the same manner, 3 mol/l sodium acetate solution (pH 5.2) and isopropanol are added to the aqueous layer at 1/10 times volume and 2 times volume, of the aqueous layer, respectively, followed by gently stirring to precipitate the genome DNA.
[0190] The genome bNk is dissolved again in a buffer containing 0.01 to 0.04 ng/ml RNase. As an example of the buffer, TR buffer (10 mmol/l Tris hydrochloride, 1 mol/l EDTA, pH 8.0) can be used. After dissolving, the resultant solution is maintained at 25 to 40° C. for 20 to 50 minutes and then extracted successively with phenol, phenol/chloroform and chloroform as in the above case.
[0191] After the extraction, isopropanol precipitation is carried out and the resulting DNA precipitate is washed with 70% ethanol, followed by air drying, and then dissolved in TE buffer to obtain a genome DNA solution.
[0192] (2) Production of shotgun library
[0193] A method for produce a genome DNA library using the genome DNA of the coryneform bacteria prepared in the above (1) include a method described in Molecular Cloning, A laboratory Manual, Second Edition (1989) (hereinafter referred to as “Molecular Cloning, 2nd ed.”). In particular, the following method can be exemplified to prepare a genome DNA library appropriately usable in determining the full nucleotide sequence by the shotgun method.
[0194] To 0.01 mg of the genome DNA of the coryneform bacteria prepared in the above (1), a buffer, such as TE buffer or the like, is added to give a total volume of 0.4 ml. Then, the genome DNA is digested into fragments of 1 to 10 kb with a sonicator (Yamato Powersonic Model SO). The treatment with the sonicator is performed at an output of 20 continuously for 5 seconds.
[0195] The resulting genome DN fragments are blunt-ended using DNA blunting kit (manufactured by Takara Shuzo) or the like.
[0196] The blunt-ended genome fragments are fractionated by agarose gel or polyacrylamide gel electrophoresis and genome fragments of 1 to 2 kb are cut out from the gel.
[0197] To the gel, 0.2 to 0.5 ml of a buffer for eluting DNA, such as MG elution buffer (0.5 mol/l amonium acetate, 10 mmol/l magnesium acetate, 1 mmol/l EDTA, 0.1% SDS) or the like, is added, followed by shaking at 25 to 40° C. overnight to elute DNA.
[0198] The resulting DNA eluate is treated with phenol/chloroform and then precipitated with ethanol to obtain a genome library insert.
[0199] This insert is ligated into a suitable vector, such as pUC18 SmaI/BAP (manufactured by Amersham Pharmacia Biotech) or the like, using T4 ligase (manufactured by Takara Shuzo) or the like. The ligation can he carried out by allowing a mixture to stand at 10 to 20° C. for 20 to 50 hours.
[0200] The resulting ligation product is precipitated with ethanol and dissolved in 5 to 20 &mgr;l of TE buffer.
[0201] Escherichia coli is transformed in accordance with a conventional method using 0.5 to 2 &mgr;l of the ligation solution. Examples of the transformation method include the electroporation method using ELECTRO MAX DH10B (manufactured by Life Technologies) for Escherichia coli. The electroporation method can be carried out under the conditions as described in the manufacturer's instructions.
[0202] The transformed Escherichia coli is spread on a suitable selection medium containing agar, for example, LB plate medium containing 10 to 100 mg/l ampicillin (LB medium (10 g/l bactotrypton, 5 g/l yeast extract, 10 g/l sodium chloride, pH 7.0) containing 1.6% of agar) when pUC18 is used as the cloning vector, and cultured therein.
[0203] The transformant can be obtained as colonies formed on the plate medium. In this step, it is possible to select the transformant having the recombinant DNA containing the genome DNA as white colonies by adding X-gal and IPTG (isopropyl-&bgr;-thiogalactopyranoside) to the plate medium.
[0204] The transformant is allowed to stand for culturing in a 96-well titer plate to which 0.05 ml of the LB medium containing 0.1 mg/ml of ampicillin has been added in each well. The resulting culture can be used in an experiment of (4) described below. Also, the culture solution can be stored at −80° C. by adding 0.05 ml per well of the LB medium containing 20% glycerol to the culture solution, followed by mixing, and the stored culture solution can be used at any time.
[0205] (3) Production of cosmid library
[0206] The genome DNA (0.l mg) of the coryneform bacteria prepared in the above (1) is partially digested with a restriction enzyme, such as Sau3AI or the like, and then ultracentrifuged (26,000 rpm, 18 hours, 20° C.) under a 10 to 40% sucrose density gradient using a 10% sucrose buffer (1 mol/l NaCl, 20 mmol/l Tris hydrochloride, 5 mnol/l EDTA, 10% sucrose, pH 8.0) and a 40% sucrose buffer (elevating the concentration of the 10% sucrose buffer to 40%).
[0207] After the centrifugation, the thus separated solution is fractionated into tubes in 1 ml per each tube. After confirming the DNA fragment size of each fraction by agarose gel electrophoresis, a fraction rich in DNA fragments of about 40 kb is precipitated with ethanol.
[0208] The resulting DNA fragment is ligated to a cosmid vector having a cohesive end which can be ligated to the fragment. When the genome DNA is partially digested with Sau3AI, the partially digested product can be ligated to, for example, the BamRI site of superCos1 (manufactured by Stratagene) in accordance with the manufacture's instructions.
[0209] The resulting ligation product is packaged using a packaging extract which can be prepared by a method described in Molecular Cloning, 2nd ed. and then used in transforming Escherichia coli. More specifically, the ligation product is packaged using, for example, a commercially available packaging extract, Gigapack III Gold Packaging Extract (manufactured by Stratagene) in accordance with the manufacture's instructions and then introduced into Escherichia coli XL-1-BlueMR (manufactured by Stratagene) or the like.
[0210] The thus transformed Escherichia coli is spread on an LB plate medium containing ampicillin, and cultured therein.
[0211] The transformant can be obtained as colonies formed on the plate medium.
[0212] The transformant is subjected to standing culture in a 96-well titer plate to which 0.05 ml of the LB medium containing 0.1 mg/ml ampicillin has been added.
[0213] The resulting culture can be employed in an experiment of (4) described below. Also, the culture solution can be stored at −80° C. by adding 0.05 ml per well of the LB medium containing 20% glycerol to the culture solution, followed by mixing, and the stored culture solution can be used at any time.
[0214] (4) Determination of nucleotide sequence
[0215] (4-1) Preparation of template
[0216] The full nucleotide sequence of genome DNA of coryneform bacteria can be determined basically according to the whole genome shotgun method (Science, 269:496-512 (1995))
[0217] The template used in the whole genome shotgun method can be prepared by PCR using the library prepared in the above (2) (DNA Research, 5:1-9 (1998)).
[0218] Specifically, the template can be prepared as follows.
[0219] The clone derived from the whole genome shotgun library is inoculated by using a replicator (manufactured by CENETIX) into each well of a 96-well plate to which 0.08 ml per well of the LB medium containing 0.1 mg/ml ampicillin has been added, followed by stationarily culturing at 37° C. overnight.
[0220] Next, the culture solution is transported, using a copy plate (manufactured by Tokken), into each well of a 96-well reaction plate (manufactured by PE Riosystems) to which 0.025 ml per well of a PCR reaction solution has been added using TaKaRa Ex Taq (manufactured by Takara Shuzo). Then, PCP is carried out in accordance with the protocol by Makino et al. (DNA Research, 5:1-9 (1998)) using GeneAmp PCR System 9700 (manufactured by PE Biosystems) to amplify the inserted fragments.
[0221] The excessive primers and nucleotides are eliminated using a kit for purifying a PCR product, and the product is used as the template in the sequencing reaction.
[0222] It is also possible to determine the nucleotide sequence using a double-stranded DNA plasmid as a template.
[0223] The double-stranded DNA plasmid used as the template can be obtained by the following method.
[0224] The clone derived from the whole genome shotgun library is inoculated into each well of a 24- or 96-well plate to which 1.5 ml per well of a 2×YT medium (16 g/l bactotrypton, 10 g/l yeast extract, 5 g/l sodium chloride, pH 7.0) containing 0.05 mg/ml ampicillin has been added, followed by culturing under shaking at 37° C. overnight.
[0225] The double-stranded DNA plasmid can be prepared from the culture solution using an automatic plasmid preparing machine K AO PI-SO (manufactured by Kurabo Industries), a multiscreen (manufactured by Millipore) or the like, according to each protocol.
[0226] To purify the plasmid, Biomek 2000 manufactured by Beckman Coulter and the like can be used.
[0227] The resulting purified double-stranded DNA plasmad is dissolved in water to give a concentration of about 0.1 mg/ml. Then, it can be used as the template in sequencing.
[0228] (4-2) Sequencing reaction
[0229] The sequencing reaction can be carried out according to a commercially available sequence kit or the like. A specific method is exemplified below.
[0230] To 6 &mgr;l of a solution of ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (manufactured by PE Biosystems), 1 to 2 pmol of an M13 regular direction primer (Ml3-21) or an M13 reverse direction primer (M13REV) (DNA Research, 5:1-9 (1998)) and 50 to 200 ng of the template prepared in the above (4-1) (the PCR product or plasmid) to give 10 gl of a sequencing reaction solution.
[0231] A dye terminator sequencing reaction (35 to 55 cycles) is carried out using this reaction solution and Gent PCR System 9700 (manufactured by PE Biosystems) or the like. The cycle parameter can be determined in accordance with a commercially available kit, for example, the manufacturer's instructions attached with ABI PRISM Big Dye Terminator Cycle Sequencing Ready Reaction Kit.
[0232] The sample can be purified using a commercially available product, such as Multi Screen HV plate (manufactured by Millipore) or the like, according to the manufacture's instructions.
[0233] The thus purified reaction product is precipitated with ethanol, dried and then used for the analysis. The dried reaction product can be stored in the dark at −30° C. and the stored reaction product can be used at any time.
[0234] The dried reaction product can be analyzed using a commercially available sequencer and an analyzer according to the manufacture's instructions
[0235] Examples of the commercially available sequencer include ABI PRISM 377 DNA Sequencer (manufactured by PP Biosystems). Example of the analyzer include ABI PRISM 3700 DNA Analyzer (manufactured by PE Biosystems).
[0236] (5) Assembly
[0237] A software, such as phred (The University of Washington) or the like, can be used as base call for use in analyzing the sequence information obtained in the above (4). A software, such as Cross_Match (The University of Washington) or SPS Cross_Match (manufactured by Southwest Parallel Software) or the like, can be used to mask the vector sequence information.
[0238] For the assembly, a software, such as phrap (The University of Washington), SPS phrap (manufactured by Southwest Parallel Software) or the like, can be used.
[0239] In the above, analysis and output of the results thereof, a computer such as UNIX, PC, Macintosh, and the like can be used.
[0240] Contig obtained by the assembly can be analyzed using a graphical editor such as consed (The University of Washington) or the like.
[0241] It is also possible to perform a series of the operations from the base call to the assembly in a lump using a script phredPhrap attached to the consed.
[0242] As used herein, software will be understood to also be referred to as a comparator.
[0243] (6) Determination of nucleotide sequence in gap part
[0244] Each of the cosmids in the cosmid library constructed in the above (3) is prepared in the same manner as in the preparation of the double-stranded DNA plasmid described in the above (4-1). The nucleotide sequence at the end of the insert fragment of the cosmid is determined using a commercially available kit, such as ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (manufactured by PE Biosystems) according to the manufacture's instructions.
[0245] About 800 cosmid clones are sequenced at both ends of the inserted fragment to detect a nucleotide sequence in the contig derived from the shotgun sequencing obtained in (5) which is coincident with the sequence. Thus, the chain linkage between respective cosmid clones and respective contigs are clarified, and mutual alignment is carried out. Furthermore, the results are compared with known physical maps to map the cosmids and the contigs. In case of Corynebacterium glutamicum ATCC 13032, a physical map of Mol. Gen. Genet., 252:255-265 (1996) can be used.
[0246] The sequence in the region which cannot be covered with the contigs (gap part) can be determined by the following method.
[0247] Clones containing sequences positioned at the ends of the contigs are selected. Among these, a clone wherein only one end of the inserted fragment has been determined is selected and the sequence at the opposite end of the inserted fragment is determined.
[0248] A shotgun library clone or a cosmid clone derived therefrom containing the sequences at the respective ends of the inserted fragments in the two contigs is identified and the full nucleotide sequence of the inserted fragment of the clone is determined.
[0249] According to this method, the nucleotide sequence of the gap part can be determined.
[0250] When no shotgun library clone or cosmid clone covering the gap part is available, primers complementary to the end sequences of the two different contigs are prepared and the DNA fragment in the gap part is amplified. Then, sequencing is performed by the primer walking method using the amplified DNA fragment as a template or by the shotgun method in which the sequence of a shotgun clone prepared from the amplified DNA fragment is determined. Thus, the nucleotide sequence of the above-described region can be determined.
[0251] In a region showing a low sequence accuracy, primers are synthesized using AUTOFINISH function and NAVIGATING function of consed (The University of Washington), and the sequence is determined by the primer walking method to improve the sequence accuracy.
[0252] Examples of the thus determined nucleotide sequence of the full genome include the full nucleotide sequence of genome of Coryebacterium glutamicum ATCC 13032 represented by SEQ ID NO:1.
[0253] (7) Determination of nucleotide sequence of microorganism genome DNA using the nucleotide sequence represented by SEQ ID NO:1
[0254] A nucleotide sequence of a polynucleotide having a homology of 80% or more with the full nucleotida sequence of Corynebacterium glutamicum ATCC 13032 represented by SEQ ID NO:1 as determined above can also be determined using the nucleotide sequence represented by SEQ ID NO:1, and the polynucleotide having at nucleotide sequence having a homology of 80% or more with the nucleotide sequence represented by SEQ ID NO:1 of the present invention is within the scope of the present invention. The term “polynucleotide having a nucleotide sequence having a homology of 80% or more with the nucleotide sequence represented by SEQ ID NO:1 of the present invention” is a polynucleotide in which a full nucleotide sequence of the chromosome DNA can be determined using as a primer an oligonucleotide composed of continuous 5 to 50 nucleotides in the nucleotide sequence represented by SEQ ID NO:1, for example, according to PCR using the chromosome DNA as a template A particularly preferred primer in determination of the full nucleotide sequence is an oligonucleotide having nucleotide sequences which are positioned at the interval of about 300 to 500 bp, and among such oligonucleotides, an oligonucleotide having a nucleotide sequence selected from DNAs encoding a protein relating to a main metabolic pathway is particularly preferred. The polynucleotide in which the full nucleotide seuence of the chromosome DNA can be determined using the oligonucleotide includes polynucleotides constituting a chromosome DNA derived from a microorganism belonging to coryneform bacteria. Such a polynucleotide is preferably a polynucleotide constituting chromosome DNA derived from a microorganism belonging to the genus Corynebacterium, more preferably a polynucleotide constituting a chromosome DNA of Corynebacterium glutamicum.
[0255] 2. Identification of ORF (open reading frame) and expression regulatory fragment and determination of the function of ORF
[0256] Based on the full nucleotide sequence data of the genome derived from coryneform bacteria determined in the above item 1, an ORF and an expression modulating fragment can be identified. Furthermore, the function of the thus determined ORF can be determined.
[0257] The ORF means a continuous region in the nucleotide sequence of mRNA which can be translated as an amino acid sequence to mature to a protein. A region of the DNA coding for the ORF of mRNA is also called ORF.
[0258] The expression modulating fragment (hereinafter referred to as “EMF”) is used herein to define a series of polynucleotide fragments which modulate the expression of the ORF or another sequence ligated operatably thereto. The expression “modulate the expression of a sequence ligated operatably” is used herein to refer to changes in the expression of a sequence due to the presence of the EMF. Examples of the EMF include a promoter, an operator, an enhancer, a silencer, a ribosome-binding sequence, a transcriptional termination sequence, and the like. In coryneform bacteria, an EMF is usually present in an intergenic segment (a fragment positioned between two genes; about 10 to 200 nucleotides in length). Accordingly, an EMF is frequently present in an intergenic segment of 10 nucleotides or longer. It is also possible to determine or discover the presence of an EMF by using known EMF sequences as a target sequence or a target structural motif (or a target motif) using an appropriate software or comparator, such as FASTA (Proc. Natl. Acad. Sci. USA, 85:2444-48 (1988)), BLAST (J. Mbl. Biol., 215:403-410 (1990)) or the like. Also, it can be identified and evaluated using a known EMF-capturing vector (for example, pKK232-8; manufactured by Amersham Pharmacia Biotech).
[0259] The term “target sequence” is used herein to refer to a nucleotide sequence composed of 6 or more nucleotides, an amino acid sequence composed of 2 or more amino acids, or a nucleotide sequence encoding this amino acid sequence composed of 2 or more amino acids. A longer target sequence appears at random in a data base at the lower possibility. The target sequence is preferably about 10 to 100 amino acid residues or about 30 to 300 nucleotide residues.
[0260] The term “target structural motif” or “target motif” is used herein to refer to a sequence or a combination of sequences selected optionally and reasonably. Such a motif is selected on the basis of the three-dimensional structure formed by the folding of a polypeptide by means known to one of ordinary skill in the art. Various motives are known.
[0261] Examples of the target motif of a polypeptide include, but are not limited to, an enzyme activity site, a protein-protein interaction site, a signal sequence, and the like. Examples of the target motif of a nucleic acid include a promoter sequence, a transcriptional regulatory factor binding sequence, a hair pin structure, and the like.
[0262] Examples of highly useful EMF include a high-expression promoter, an inducible-expression promoter, and the like. Such an EMF can be obtained by positionally determining the nucleotide sequence of a gene which is known or expected as achieving high expression (for example, ribosomal RNA gene:GenBank Accession No. M16175 or Z46753) or a gene showing a desired induction pattern (for example, isocitrate lyase gene induced by acetic acid:Japanese Published Unexamined Patent Application No. 56782/93) via the alignment with the full genome nucleotide sequence determined in the above item 1, and isolating the genome fragment in the upstream part (usually 200 to 500 nucleotides from the translation initiation site). It is also possible to obtain a highly useful EMF by selecting an EMF showing a high expression efficiency or a desired induction pattern from among promoters captured by the EMF-capturing vector as described above.
[0263] The ORF can be identified by extracting characteristics common to individual ORFs, constructing a general model based on these characteristics, and measuring the conformity of the subject sequence with the model. In the identification, a software, such as GeneMark (Nuc. Acids. Res., 22:4756-67 (1994):manufactured by GenePro)), GeneMark.hrm (manufactured by GenePro), GeneHacker (Protein, Nucleic Acid and Enzyme, 42:3001-07 (1997)), Glimmer (Nuc. Acids. Res., 26:544-548 (1998):manufactured by The Institute of Genomic Research), or the like, can be used. In using the software, the default (initial setting) parameters are usually used, though the parameters can be optionally changed.
[0264] In the above-described comparisons, a computer, such as UNIX, PC, Macintosh, or the like, can be used.
[0265] Examples of the ORF determined by the method of the present invention include ORFs having the nucleotide sequences represented by SEQ ID NOS:2 to 3501 present in the genome of Corynebacterium glutamicum as represented by SEQ ID NO:1. In these ORFs, polypeptides having the amino acid sequences represented by SEQ ID NOS.,3502 to 7001 are encoded.
[0266] The function of an ORF can be determined by comparing the identified amino acid sequence of the ORF with known homologous sequences using a homology searching software or comparator, such as BLAST, FAST, Smith & Waterman (Meth. Enzym., 164:765 (1988)) or the like on an amino acid data base, such as Swith-Prot, PIR, GenBank-nr-aa, GenPept constituted by protein-encoding domains derived from GenBank data base, OWL or the like.
[0267] Furthermore, by the homology searching, the identity and similarity with the amino acid sequences of known proteins can also be analyzed.
[0268] With respect of the term “identity” used herein, where two polypeptides each having 10 amino acids are different in the positions of 3 amino acids, these polypeptides have an identity of 70% with each other. In case wherein one of the different 3 amino acids is analogue (for example, leonine and isoleucine), these polypeptides have a similarity of 80%.
[0269] As a specific example, Table 1 shows the registration numbers in known data bases of sequences which are judged as having the highest similarity with the nucleotide sequence of the ORF derived from Corynebacterium glutamicum ATCC 13032, genes of these sequences, functions of these genes, and identities thereof compared with known amino acid translation sequences.
[0270] Thus, a great number of novel genes derived from coryneform bacteria can be identified by determining the full nucleotide sequence of the genome derived from coryneform bacterium by the means of the present invention. Moreover, the function of the proteins encoded by these genes can be determined. Since coryneform bacteria are industrially highly useful microorganisms, many of the identified genes are industrially useful.
[0271] Moreover, the characteristics of respective microorganisms can be clarified by classifying the functions thus determined. As a result, valuable information in breeding is obtained.
[0272] Furthermore, from the ORF information derived from coryneform bacteria, the ORF corresponding to the :microorganism is prepared and obtained according to the general method as disclosed in Molecular Cloning, 2nd ed. or the like. Specifically, an oligonucleotide having a nucleotide sequence adjacent to the ORF is synthesized, and the ORF can be isolated and obtained using the oligonucleotide as a primer and a chromosome DNA derived from coryneform bacteria as a template according to the general PCR cloning technique. Thus obtained ORF sequences include polynucleotides comprising the nucleotide sequence represented by any one of SEQ ID NOS:2 to 3501.
[0273] The ORF or primer can be prepared using a polypeptide synthesizer based on the above sequence information.
[0274] Examples of the polynucleotide of the present invention include a polynucleotide containing the nucleotide sequence of the ORBF obtained in the above, and a polynucleotide which hybridizes with the polynucleotide under stringent conditions.
[0275] The polynucleotide of the present invention can be a single-stranded DNA, a double-stranded DNA and a single-stranded RNA, though it is not limited thereto.
[0276] The polynucleotide which hybridizes with the polynucleotide containing the nucleotide sequence of the ORP obtained in the above under stringent conditions includes a degenerated mutant of the ORF. A degenerated mutant is a polynucleotide fragment having a nucleotide sequence which is different from the sequence of the ORF of the present invention which encodes the same amino acid sequence by degeneracy of a gene code.
[0277] Specific examples include a polynucleotide comprising the nucleotide sequence represented by any one of SEQ ID NOS:2 to 3431, and a polynucleotide which hybridizes with the polynucleotide under stringent conditions.
[0278] A polynucleotide which hybridizes under stringent conditions is a polynucleotide obtained by colony hybridization, plaque hybridization, Southern blot hybridization or the like using, as a probe, the polynucleotide having the nucleotide sequence of the ORF identified in the above. Specific examples include a polynucleotide which can be identified by carrying out hybridization at 65° C. in the presence of 0.7-1.0 M NaCl using a filter on which a polynucleotide prepared from colonies or plaques is immobilized, and then washing the filter with 0.1× to 2× SSC solution (the composition of 1× SSC contains 150 mM sodium chloride and 15 mM sodium citrate) at 65° C.
[0279] The hybridization can be carried out in accordance with known methods described in, for example, Molecular Cloning, 2nd ed., Current Protocols in Molecular Biology, DNA Cloning 1:Core Techniques, A Practical Approach, Second Edition, Oxford University (1995) or the like. Specific examples of the polynucleotide which can be hybridized include a DNA having a homology of 60% or more, preferably 80% or more, and particularly preferably 95% or more, with the nucleotide sequence represented by any one of SEQ ID NO:2 to 3431 when calculated using default (initial setting) parameters of a homology searching software, such as BLAST, FASTA, Smith-Waterman or the like.
[0280] Also, the polynucleotide of the present invention includes a polynucleotide encoding a polypeptide comprising the amino acid sequence represented by any one of SEQ ID NOS:3502 to 6931 and a polynucleotide which hybridizes with the polynucleotide under stringent conditions.
[0281] Furthermore, the polynucleotide of the present invention includes a polynucleotide which is present in the 51 upstream or 3′ downstream region of a polynucleotide comprising the nucleotide sequence of any one of SEQ ID NOS:2 to 3431 in a polynucleotide comprising the nucleotide sequence represented by SEQ ID NO:1, and has an activity of regulating an expression of a polypeptide encoded by the polynucleotide. Specific examples of the polynucleotide having an activity of regulating an expression of a polypeptide encoded by the polynucleotide includes a polynucleotide encoding the above described EMF, such as a promoter, an operator, an enhancer, a silencer, a ribosome-binding sequence, a transcriptional termination sequence, and the like.
[0282] The primer used for obtaining the ORF according to the above PCR cloning technique includes an oligonucleotide comprising a sequence which is the same as a sequence of 10 to 200 continuous nucleotides in the nucleotide sequence of the ORF and an adjacent region or an oligonucleotide comprising a sequence which is complementary to the oligonucleotide. Specific examples include an oligonucleotide comprising a sequence which is the same as a sequence of 10 to 200 continuous nucleotides of the nucleotide sequence represented by any one of SEQ ID NOS:1 to 3431, and an oligonucleotide comprising a sequence complementary to the oligonucleotide comprising a sequence of at least 10 to 20 continuous nucleotide of any one of SEQ ID NOS:1 to 3431. When the primers are used as a sense primer and an antisense primer, the above-described oligonucleotides in which melting temperature (Tm) and the number of nucleotides are not significantly different from each other are preferred.
[0283] The oligonucleotide of the present invention includes an oligonucleotide comprising a sequence which is the same as 10 to 200 continuous nucleotides of the nucleotide sequence represented by any one of SEQ ID NOS:1 to 3431 or an oligonucleotide comprising a sequence complementary to the oligonucleotide.
[0284] Also, analogues of these oligonucleotides (hereinafter also referred to as “analogous oligonucleotides”) are also provided by the present invention and are useful in the methods described herein.
[0285] Examples of the analogous oligonucleotides include analogous oligonucleotides in which a phosphodiester bond in an oligonucleotide is converted to a phosphorothioate bond, analogous oligonucleotides in which a phosphodiester bond in an oligonucleotide is converted to an N3′-P5′ phosphoamidate bond, analogous oligonucleotides in which ribosome and a phosphodiester bond in an oligonucleotide is converted to a peptide nucleic acid bond, analogous oligonucleotides in which uric in an oligonucleotide is replaced with C-5 propynyluracil, analogous oligonucleotides in which uric in an oligonucleotide is replaced with C-5 thiazoluracil, analogous oligonucleotides in which cytosine in an oligonucleotide is replaced with C-5 propynylcytosine, analogous oligonucleotides in which cytosine in an oligonucleotide is replaced with phenoxazine-modified cytosine, analogous oligonucleotides in which ribose in an oligonucleotide is replaced with 2′-O-propylribose, analogous oligonucleotides in which ribose in an oligonucleotide is replaced with 2′-methoxyethoxyribose, and the like (Cell Engineering, 16:1463 (1997)).
[0286] The above oligonucleotides and analogous oligonucleotides of the present invention can be used as probes for hybridization and antisense nucleic acids described below in addition to as primers.
[0287] Examples of a primer for the antisense nucleic acid techniques known in the art include an oligonucleotide which hybridizes the oligonucleotide of the present invention under stringent conditions and has an activity regulating expression of the polypeptide encoded by the polynucleotide, in addition to the above oligonucleotide.
[0288] 3. Determination of isozymes
[0289] Many mutants of coryneform bacteria which are useful in the production of useful substances, such as amino acids, nucleic acids, vitamins, saccharides, organic acids, and the like, are obtained by the present invention.
[0290] However, since the gene sequence data of the microorganism has been, to date, insufficient, useful mutants have been obtained by mutagenic techniques using a mutagen, such as nitrosoguanidine (NTG) or the like.
[0291] Although genes can be mutated randomly by the mutagenic method using the above-described mutagen, all genes encoding respective isozymes having similar properties relating to the metabolism of intermediates cannot be mutated. In the mutagenic method using a mutagen, genes are mutated randomly. Accordingly, harmful mutations worsening culture characteristics, such as delay in growth, accelerated foaming, and the like, might be imparted at a great frequency, in a random manner.
[0292] However, if gene sequence information is available, such as is provided by the present invention, it is possible to mutate all of the genes encoding target isozymes. In this case, harmful mutations may be avoided and the target mutation can be incorporated.
[0293] Namely, an accurate number and sequence information of the target isozymes in coryneform bacteria can be obtained based on the ORF data obtained in the above item 2. By using the sequence information, all of the target isozyme genes can be mutated into genes having the desired properties by, for example, the site-specific mutagenesis method described in Molecular Cloning, 2nd ed. to obtain useful mutants having elevated productivity of useful substances.
[0294] 4. Clarification or determination of biosynthesis pathway and signal transmission pathway
[0295] Attempts have been made to elucidate biosynthesis pathways and signal transmission pathways in a number of organisms, and many findings have been reported. However, there are many unknown aspects of coryneform bacteria since a number of genes have not been identified so far.
[0296] These unknown points can be clarified by the following method.
[0297] The functional information of ORF derived from coryneform bacteria as identified by the method of above item 2 is arranged. The term “arranged” means that the ORF is classified based on the biosynthesis pathway of a substance or the signal transmission pathway to which the ORF belongs using known information according to the functional information. Next, the arranged ORF sequence information is compared with enzymes on the biosynthesis pathways or signal transmission pathways of other known organisms. The resulting information is combined with known data on coryneform bacteria. Thus, the biosynthesis pathways and signal transmission pathways in coryneform bacteria, which have been unknown so far, can be determined.
[0298] As a result that these pathways which have been unknown or unclear hitherto are clarified, a useful mutant for producing a target useful substance can be efficiently obtained.
[0299] When the thus clarified pathway is judged as important in the synthesis of a useful product, a useful mutant can be obtained by selecting a mutant wherein this pathway has been strengthened. Also, when the thus clarified pathway is judged as not important in the biosynthesis of the target useful product, a useful mutant can be obtained by selecting a mutant wherein the utilization frequency of this pathway is lowered.
[0300] 5. Clarification or determination of useful mutation point
[0301] Many useful mutants of coryneform bacteria which are suitable for the production of useful substances, such as amino acids, nucleic acids, vitamins, saccharides, organic acids, and the like, have been obtained. However, it is hardly known which mutation point is imparted to a gene to improve the productivity.
[0302] However, mutation points contained in production strains can be identified by comparing desired sequences of the genome DNA of the production strains obtained from coryneform bacteria by the mutagenic technique with the nucleotide sequences of the corresponding genome DNA and ORF derived from coryneform bacteria determined by the methods of the above items 1 and 2 and analyzing them
[0303] Moreover, effective mutation points contributing to the production can be easily specified from among these mutation points on the basis of known information relating to the metabolic pathways, the metabolic regulatory mechanisms, the structure activity correlation of enzymes, and the like.
[0304] When any efficient mutation can be hardly specified based on known data, the mutation points thus identified can be introduced into a wild strain of coryneform bacteria or a production strain free of the mutation. Then, it is examined whether or not any positive effect can be achieved on the production.
[0305] For example, by comparing the nucleotide sequence of homoserine dehydrogenase gene hom of a lysine-producing B-6 strain of Corynebacterium glutamicum (Appl. Microbiol. Biotechnol., 32:269-273 (1989)) with the nucleotide sequence corresponding to the genome of Corynebacterium qiutamicum ATCC 13032 according to the present invention, a mutation of amino acid replacement in which valine at the 59-position is replaced with alanine (Val59Ala) was identified. A strain obtained by introducing this mutation into the ATCC 13032 strain by the gene replacement method can produce lysine, which indicates that this mutation is an effective mutation contributing to the production of lysine.
[0306] Similarly, by comparing the nucleotide sequence of private carboxylase gene pyc of the B-6 strain with the nucleotide sequence corresponding to the ATCC 13032 genome, a mutation of amino acid replacement in which proline at the 458-position was replaced with serine (Pro458Ser) was identified. A strain obtained by introducing this mutation into a lysine-producing strain of No. 58 (FERM BP-7134) of Corynebacterium glutamicum free of this mutation shows an improved lysine productivity in comparison with the No. 58 strain, which indicates that this mutation is an effective mutation contributing to the production of lysine.
[0307] In addition, a mutation Ala2l3Thr in glucose-6-phosphate dehydrogenase was specified as an effective mutation relating to the production of lysine by detecting glucose-6-phosphate dehydrogenase gene zwf of the B-6 strain.
[0308] Furthermore, the lysine-productivity of Corynebacterium glutamicum was improved by replacing the base at the 932-position of aspartokinase gene lysC of the Corynebacterium glutamicum ATCC 13032 genome with cytosine to thereby replace threonine at the 311-position by isoleucine, which indicates that this mutation is an effective mutation contributing to the production of lysine.
[0309] Also, as another method to examine whether or not the identified mutation point is an effective mutation, there is a method in which the mutation possessed by the lysine-producing strain is returned to the sequence of a wild type strain by the gene replacement method and whether or not it has a negative influence on the lysine productivity. For example, when the amino acid replacement mutation Val59Ala possessed by hom of the lysine-producing B-6 strain was returned to a wild type amino acid sequence, the lysine productivity was lowered in comparison with the B-6 strain. Thus, it was found that this mutation is an effective mutation contributing to the production of lysine.
[0310] Effective mutation points can be more efficiently and comprehensively extracted by combining, if needed, the DNA array analysis or proteome analysis described below.
[0311] 6. Method of breeding industrially advantageous production strain
[0312] It has been a general practice to construct production strains, which are used industrially in the fermentation production of the target useful substances, such as amino acids, nucleic acids, vitamins, saccharides, organic acids, and the like, by repeating mutagenesis and breeding based on random mutagenesis using mutagens, such as NTG or the like, and screening.
[0313] In recent years, many examples of improved production strains have been made through the use of recombinant DNA techniques. In breeding, however, most of the parent production strains to be improved are mutants obtained by a conventional mutagenic procedure (W. Leuchtenberger, Amino Acids—Technical Production and Use. In:Roehr (ed) Biotechnology, second edition, vol. 6, products of primary metabolism. VCH Verlagsgesellschaft mbH, Weinheim, P 465 (1996)).
[0314] Although mutagenesis methods have largely contributed to the progress of the fermentation industry, they suffer from a serious problem of multiple, random introduction of mutations into every part of the chromosome. Since many mutations are accumulated in a single chromosome each time a strain is improved, a production strain obtained by the random mutation and selecting is generally inferior in properties (for example, showing poor growth, delayed consumption of saccharides, and poor resistance to stresses such as temperature and oxygen) to a wild type strain, which brings about troubles such as failing to establish a sufficiently elevated productivity, being frequently contaminated with miscellaneous bacteria, requiring troublesome procedures in culture maintenance, and the like, and, in its turn, elevating the production cost in practice. In addition, the improvement in the productivity is based on random mutations and thus the mechanism thereof is unclear. Therefore, it is very difficult to plan a rational breeding strategy for the subsequent improvement in the productivity.
[0315] According to the present invention, effective mutation points contributing to the production can be efficiently specified from among many mutation points accumulated in the chromosome of a production strain which has been bred from coryneform bacteria and, therefore, a novel breeding method of assembling these effective mutations in the coryneform bacteria can be established. Thus, a useful production strain can be reconstructed. It is also possible to construct a useful production strain from a wild type strain.
[0316] Specifically, a useful mutant can be constructed in the following manner.
[0317] One of the mutation points is incorporated into a wild type strain of coryneform bacteria. Then, it is examined whether or not a positive effect is established on the production. When a positive effect is obtained, the mutation point is saved. When no effect is obtained, the mutation point is removed. Subsequently, only a strain having the effective mutation point is used as the parent strain, and the same procedure is repeated. In general, the effectiveness of a mutation positioned upstream cannot be clearly evaluated in some cases when there is a rate-determining point in the downstream of a biosynthesis pathway. It is therefore preferred to successively evaluate mutation points upward from downstream.
[0318] By reconstituting effective mutations by the method as described above in a wild type strain or a strain which has a high growth speed or the same ability to consume saccharides as the wild type strain, it is possible to construct an industrially advantageous strain which is free of troubles in the previous methods as described above and to conduct fermentation production using such strains within a short time or at a higher temperature.
[0319] For example, a lysine-producing mutant B-6 (Appl. Microbiol. Biotechnol., 32:262-273 (1989)), which is obtained by multiple rounds of random mutagenesis from a wild type strain Corynebacterium glutamicum ATCC 13032, enables lysine fermentation to be performed at a temperature between 30 and 34° C. but shows lowered growth and lysine productivity at a temperature exceeding 34° C. Therefore, the fermentation temperature should be maintained at 34° C. or lower. In contrast thereto, the production strain described in the above item 5, which is obtained by reconstituting effective mutations relating to lysine production, can achieve a productivity at 40 to 42° C. equal or superior to the result obtained by culturing at 30 to 34° C. Therefore, this strain is industrially advantageous since it can save the load of cooling during the fermentation.
[0320] When culture should be carried out at a high temperature exceeding 43° C., a production strain capable of conducting fermentation production at a high temperature exceeding 43° C. can be obtained by reconstituting useful mutations in a microorganism belonging to the genus Corynebacterium which can grow at high temperature exceeding 43° C. Examples of the microorganism capable of growing at a high temperature exceeding 43° C. include Corynebacterium thermoaminogenes, such as Corynebacterium thexmoaminogenes FERM 9244, PERM 9245, FERM 9246 and FERM 9247
[0321] A strain having a further improved productivity of the target product can be obtained using the thus reconstructed strain as the parent strain and further breeding it using the conventional mutagenesis method, the gene amplification method, the gene replacement method using the recombinant DNA technique, the transduction method or the cell fusion method. Accordingly, the microorganism of the present invention includes, but is not limited to, a mutant, a cell fusion strain, a transformant, a transductant or a recombinant strain constructed by using recombinant DNA techniques, so long as it is a producing strain obtained via the step of accumulating at least two effective mutations in a coryneform bacteria in the course of breeding.
[0322] When a mutation point judged as being harmful to the growth or production is specified, on the other hand, it is examined whether or not the producing strain used at present contains the mutation point. When it has the mutation, it can be returned to the wild type gene and thus a further useful production strain can be bred.
[0323] The breeding method as described above is applicable to microorganisms, other than coryneform bacteria, which have industrially advantageous properties (for example, microorganisms capable of quickly utilizing less expensive carbon sources, microorganisms capable of growing at higher temperatures).
[0324] 7. Production and utilization of polynucleotide array
[0325] (1) Production of polynucleotide array
[0326] A polynucleotide array can be produced using the polynucleotide or oligonucleotide of the present invention obtained in the above items 1 and 2.
[0327] Examples include a polynucleotide array comprising a solid support to which at least one of a polynucleotide it comprising the nucleotide sequence represented by SEQ ID NOS:2 to 3501, a polynucleotide which hybridizes with the polynucleotide under stringent conditions, and a polynucleotide comprising 10 to 200 continuous nucleotides in the nucleotide sequence of the polynucleotide is adhered; and a polynucleotide array comprising a solid support to which at least one of a polynucleotide encoding a polypeptide comprising the amino acid sequence represented by any one of SEQ ID NOS:3502 to 7001, a polynucleotide which hybridizes with the polynucleotide under stringent conditions, and a polynucleotide comprising 10 to 200 continuous bases in the nucleotide sequences of the polynucleotides is adhered.
[0328] Polynucleotide arrays of the present invention include substrates known in the art, such as a DNA chip, a DNA microarray and a DNA macroarray, and the like, and comprises a solid support and plural polynucleotides or fragments thereof which are adhered to the surface of the solid support.
[0329] Examples of the solid support include a glass plate, a nylon membrane, and the like.
[0330] The polynucleotides or fragments thereof adhered to the surface of the solid support can be adhered to the surface of the solid support using the general technique for preparing arrays. Namely, a method in which they are adhered to a chemically surface-treated solid support, for example, to which a polycation such as polylysine or the like has been adhered (Nat. Genet., 21:15-19 (1999)). The chemically surface-treated supports are commercially available and the commercially available solid product can be used as the solid support of the polynucleotide array according to the present invention.
[0331] As the polynucleotides or oligonucleotides adhered to the solid support, the polynucleotides and oligonucleotides of the present invention obtained in the above items 1 and 2 can be used.
[0332] The analysis described below can be efficiently performed by adhering the polynucleotides or oligonucleotides to the solid support at a high density, though a high fixation density is not always necessary.
[0333] Apparatus for achieving a high fixation density, such as an arrayer robot or the like, is commercially available from Takara Shuzo (GMS417 Arrayer), and the commercially available product can be used.
[0334] Also, the oligonucleotides of the present invention can be synthesized directly on the solid support by the photolithography method or the like (Nat. Genet., 21:20-24 (1999)). In this method, a linker having a protective group which can be removed by light irradiation is first adhered to a solid support, such as a slide glass or the like. Then, it is irradiated with light through a mask (a photolithograph mask) permeating light exclusively at a definite part of the adhesion part. Next, an oligonucleotide having a protective group which can be removed by light irradiation is added to the part. Thus, a ligation reaction with the nucleotide arises exclusively at the irradiated part. By repeating this procedure, oligonucleotides, each having a desired sequence, different from each other can be synthesized in respective parts. Usually, the oligonucleotides to be synthesized have a length of 10 to 30 nucleotides.
[0335] (2) Use of polynucleotide array
[0336] The following procedures (a) and (b) can be carried out using the polynucleotide array prepared in the above (1).
[0337] (a) Identification of mutation point of coryneform bacterium mutant and analysis of expression amount and expression profile of gene encoded by genome
[0338] By subjecting a gene derived from a mutant of coryneform bacteria or an examined gene to the following steps (i) to (iv), the mutation point of the gene can be identified or the expression amount and expression profile of the gene can be analyzed:
[0339] (i) producing a polynucleotide array by the method of the above (1);
[0340] (ii) incubating polynucleotides immobilized on the polynucleotide array together with the labeled gene derived from a mutant of the coryneform bacterium using the polynucleotide array produced in the above (i) under hybridization conditions;
[0341] (iii) detecting the hybridization; and
[0342] (iv) analyzing the hybridization data.
[0343] The gene derived from a mutant of coryneform bacteria or the examined gene include a gene relating to biosynthesis of at least one selected from amino acids, nucleic acids, vitamins, saccharides, organic acids, and analogues thereof.
[0344] The method will be described in detail.
[0345] A single nucleotide polymorphism (SNP) in a human region of 2,300 kb has been identified using polynucleotide arrays (Science, 280:1077-82 (1998)). In accordance with the method of identifying SNP and methods described in Science, 278:680-686 (1997); Proc. Natl. Acad. Sci. USA, 96:12833-38 (1999); Science, 284:1520-23 (1999), and the like using the polynucleotide array produced in the above (1) and a nucleic acid molecule (DNA, RNA) derived from coryneform bacteria in the method of the hybridization, a mutation point of a useful mutant, which is useful in producing an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, or the like can be identified and the gene expression amount and the expression profile thereof can be analyzed.
[0346] The nucleic acid molecule (DNA, RNA) derived from the coryneform bacteria can be obtained according to the general method described in Molecular Cloning, 2nd ed. or the like. MRNA derived from Corynebacterium glutamicum can also be obtained by the method of Bormann et al. (Molecular Microbiology, 6:317-326 (1992)) or the like.
[0347] Although ribosomal RNA (rRNA) is usually obtained in large excess in addition to the target MRNA, the analysis is not seriously disturbed thereby.
[0348] The resulting nucleic acid molecule derived from coryneform bacteria is labeled. Labeling can be carried out according to a method using a fluorescent dye, a method using a radioisotope or the like.
[0349] Specific examples include a labeling method in which psoralen-biotin is crosslinked with RNA extracted from a microorganism and, after hybridization reaction, a fluorescent dye having streptoavidin bound thereto is bound to the biotin moiety (Nat. Biotechnol., 16:45-48 (1998)); a labeling method in which a reverse transcription reaction is carried out using RNA extracted from a microorganism as a template and random primers as primers, and dUTP having a fluorescent dye (for example, Cy3, Cy5) (manufactured by Amersham Pharmacia Biotech) is incorporated into CDNA (Proc. Natl. Acad. Sci. USA, 96:12833-38 (1999)); and the like.
[0350] The labeling specificity can be improved by replacing the random primers by sequences complementary to the 3′-end of ORF (J. Bacteriol., 181:6425-40 (1999)).
[0351] In the hybridization method, the hybridization and subsequent washing can be carried out by the general method (Nat. Bioctechnol., 14:1675-80 (1996), or the like).
[0352] Subsequently, the hybridization intensity is measured depending on the hybridization amount of the nucleic acid molecule used in the labeling. Thus, the mutation point can be identified and the expression amount of the gene can be calculated.
[0353] The hybridization intensity can be measured by visualizing the fluorescent signal, radioactivity, luminescence dose, and the like, using a laser confocal microscope, a CCD camera, a radiation imaging device (for example, STORM manufactured by Amersham Pharmacia Biotech), and the like, and then quantifying the thus visualized data.
[0354] A polynucleotide array on a solid support can also be analyzed and quantified using a commercially available apparatus, such as GMS418 Array Scanner (manufactured by Takara Shuzo) or the like.
[0355] The gene expression amount can be analyzed using a commercially available software (for example, ImaGene manufactured by Takara Shuzo; Array Gauge manufactured by Fuji Photo Film; ImageQuant manufactured by Amersham Pharmacia Biotech, or the like).
[0356] A fluctuation in the expression amount of a specific gene can be monitored using a nucleic acid molecule obtained in the time course of culture as the nucleic acid molecule derived from coryneform bacteria. The culture conditions can be optimized by analyzing the fluctuation.
[0357] The expression profile of the microorganism at the total gene level (namely, which genes among a great number of genes encoded by the genome have been expressed and the expression ratio thereof) can be determined using a nucleic acid molecule having the sequences of many genes determined from the full genome sequence of the microorganism. Thus, the expression amount of the genes determined by the full genome sequence can be analyzed and, in its turn, the biological conditions of the microorganism can be recognized as the expression pattern at the full gene level.
[0358] (b) Confirmation of the presence of gene homologous to examined gene in coryneform bacteria
[0359] Whether or not a gene homologous to the examined gene, which is present in an organism other than coryneform bacteria, is present in coryneform bacteria can be detected using the polynucleotide array prepared in the above (1).
[0360] This detection can be carried out by a method in which an examined gene which is present in an organism other than coryneform bacteria is used instead of the nucleic acid molecule derived from coryneform bacteria used in the above identification/analysis method of (1).
[0361] 8. Recording medium storing full genome nucleotide sequence and ORF data and being readable by a computer and methods for using the same
[0362] The term “recording medium or storage device which is readable by a computer” means a recording medium or storage medium which can be directly readout and accessed with a computer. Examples include magnetic recording media, such as a floppy disk, a hard disk, a magnetic tape, and the like; optical recording media, such as CD-ROM, CD-R, CD-RW, DVD-ROM, DVD-RAM, DVD-RW, and the like; electric recording media, such as RAM, ROM, and the like; and hybrids in these categories (for example, magnetic/optical recording media, such as MO and the like).
[0363] Instruments for recording or inputting in or on the recording medium or instruments or devices for reading out the information in the recording medium can be appropriately selected, depending on the type of the recording medium and the access device utilized. Also, various data processing programs, software, comparator and formats are used for recording and utilizing the polynucleotide sequence information or the like of the present invention in the recording medium. The information can be expressed in the form of a binary file, a text file or an ASCII file formatted with commercially available software, for example. Moreover, software for accessing the sequence information is available and known to one of ordinary skill in the art.
[0364] Examples of the information to be recorded in the above-described medium include the full genome nucleotide sequence information of coryneform bacteria as obtained in the above item 2, the nucleotide sequence information of ORF, the amino acid sequence information encoded by the ORF, and the functional information of polynucleotides coding for the amino acid sequences.
[0365] The recording medium or storage device which is readable by a computer according to the present invention refers to a medium in which the information of the present invention has been recorded. Examples include recording media or storage devices which are readable by a computer storing the nucleotide sequence information represented by SEQ ID NOS:1 to 3501, the amino acid sequence information represented by SEQ ID NOS:3502 to 7001, the functional information of the nucleotide sequences represented by SEQ ID NOS:1 to 3501, the functional information of the amino acid sequences represented by SEQ ID NOS:3502 to 7001, and the information listed in Table 1 below and the like.
[0366] 9. System based on a computer using the recording medium of the present invention which is readable by a computer
[0367] The term “system based on a computer” as used herein refers a system composed of hardware device(s), software device(s), and data recording device(s) which are used for analyzing the data recorded in the recording medium of the present invention which is readable by a computer.
[0368] The hardware device(s) are, for example, composed of an input unit, a data recording unit, a central processing unit and an output unit collectively or individually.
[0369] By the software device(s), the data recorded in the recording medium of the present invention are searched or analyzed using the recorded data and the hardware device(s) as described herein. Specifically, the software device(s) contain at least one program which acts on or with the system in order to screen, analyze or compare biologically meaningful structures or information from the nucleotide sequences, amino acid sequences and the like recorded in the recording medium according to the present invention.
[0370] Examples of the software device(s) for identifying ORF and EMF domains include GeneMark (Nuc. Acids. Res., 22:4756-67 (1994)), GeneHacker (Protein, Nucleic Acid and Enzyme, 42:3001-07 (1997)), Glimmer (The Institute of Genomic Research; Nuc. Acids. Res., 26:544-548 (1998)) and the like. In the process of using such a software device, the default (initial setting) parameters are usually used, although the parameters can be changed, if necessary, in a manner known to one of ordinary skill in the art.
[0371] Examples of the software device(s) for identifying a genome domain or a polypeptide domain analogous to the target sequence or the target structural motif (homology searching) include FASTA, BLAST, Smith-Waterman, GenetyxMac (manufactured by Software Development), GCG Package (manufactured by Genetic Computer Group), GenCore (manufactured by Compugen), and the like. In the process of using such a software device, the default (initial setting) parameters are usually used, although the parameters can be changed, if necessary, in a manner known to one of ordinary skill in the art.
[0372] Such a recording medium storing the full genome sequence data is useful in preparing a polynucleotide array by which the expression amount of a gene encoded by the genome DNA of coryneform bacteria and the expression profile at the total gene level of the microorganism, namely, which genes among many genes encoded by the genome have been expressed and the expression ratio thereof, can be determined.
[0373] The data recording device(s) provided by the present invention are, for example, memory device(s) for recording the data recorded in the recording medium of the present invention and target sequence or target structural motif data, or the like, and a memory accessing device(s) for accessing the same.
[0374] Namely, the system based on a computer according to the present invention comprises the following:
[0375] (i) a user input device that inputs the information stored in the recording medium of the present invention, and target sequence or target structure motif information;
[0376] (ii) a data storage device for at least temporarily storing the input information;
[0377] (iii) a comparator that compares the information stored in the recording medium of the present invention with the target sequence or target structure motif information, recorded by the data storing device of (ii) for screening and analyzing nucleotide sequence information which is coincident with or analogous to the target sequence or target structure motif information; and
[0378] (iv) an output device that shows a screening or analyzing result obtained by the comparator.
[0379] This system is usable in the methods in items 2 to 5 as described above for searching and analyzing the ORP and ZMF domains, target sequence, target structural motif, etc. of a coryneform bacterium, searching homologs, searching and analyzing isozymes, determining the biosynthesis pathway and the signal transmission pathway, and identifying spots which have been found in the proteome analysis. The term “homologs” as used herein includes both of orthologs and paralogs.
[0380] 10. Production of polypeptide using ORF derived from coryneform bacteria
[0381] The polypeptide of the present invention can be produced using a polynucleotide comprising the ORF obtained in the above item 2. Specifically, the polypeptide of the present invention can be produced by expressing the polynucleotide of the present invention or a fragment thereof in a host cell, using the method described in Molecular Cloning, 2nd ed., Current Protocols in Molecular Biology, and the like, for example, according to the following method.
[0382] A DNA fragment having a suitable length containing a part encoding the polypeptide is prepared from the full length ORF sequence, if necessary.
[0383] Also, DNA in which nucleotides in a nucleotide sequence at a part encoding the polypeptide of the present invention are replaced to give a codon suitable for expression of the host cell, if necessary. The DNA is useful for efficiently producing the polypeptide of the present invention.
[0384] A recombinant vector is prepared by inserting the DNA fragment into the downstream of a promoter in a suitable expression vector.
[0385] The recombinant vector is introduced to a host cell suitable for the expression vector.
[0386] Any of bacteria, yeasts, animal cells, insect cells, plant cells, and the like can be used as the host cell so long as it can be expressed in the gene of interest.
[0387] Examples of the expression vector include those which can replicate autonomously in the above-described host cell or can be integrated into chromosome and have a promoter at such a position that the DNA encoding the polypeptide of the present invention can be transcribed.
[0388] When a procaryote cell, such as a bacterium or the like, is used as the host cell, it is preferred that the recombinant vector containing the DNA encoding the polypeptide of the present invention can replicate autonomously in the bacterium and is a recombinant vector constituted by, at least a promoter, a ribosome binding sequence, the DNA of the present invention and a transcription termination sequence. A promoter controlling gene can also be contained therewith in operable combination.
[0389] Examples of the expression vectors include a vector plasmid which is replicable in Corynebacterium glutamicum, such as pCGl (Japanese Published Unexamined Patent Application No. 134500/82), pCG2 (Japanese Published Unexamined Patent Application No. 35197/83), pCG4 (Japanese Published Unexamined Patent Application No. 183799/82), pCGll (Japanese Published Unexamined Patent Application No. 134500/82), pCG116, pCE54 and pCB101 (Japanese Published Unexamined Patent Application No. 105999/83), pCE51, pCE52 and pCE53 (Mol. Gen. Genet., 196:175-178 (1984)), and the like; a vector plasmid which is replicable in Escherichia coli, such as pET3 and pET1l (manufactured by Stratagene), PBAD, pThioHis and pTrcHis (manufactured by Invitrogen), pKK223-3 and pGEX2T (manufactured by Amersham Pharmacia Biotech), and the like; and pBTrp2, pBTacl and pBTac2 (manufactured by Boehringer Mannheim Co.), pSE280 (manufactured by Invitrogen), pGEMEX-1 (manufactured by Promega), pQE-8 (manufactured by QIAGEN), pKYP1O (Japanese Published Unexamined Patent Application No. 110600/83), pKYP200 (Agric. Biol. Chem., 48:669 (1984)), pLSAl (Agric. Biol. Chem., 53:277 (1989)), pGEL1 (Proc. Natl. Acad. Sci. USA, 82:4306 (1985)), pBluescript II SK(−) (manufactured by Stratagene), pTrs3O (prepared from Escherichia coli JM109/pTrS30 (FERM BP-5407)), pTrs32 (prepared from Escherichia coli JM109/pTrS32 (FERM BP-5408)), pGHA2 (prepared from Escherichia coli IGHA2 (FERM B-400), Japanese Published Unexamined Patent Application No. 221091/85), pGKA2 (prepared from Escherichia coli IGKA2 (FERM BP-6798), Japanese Published Unexamined Patent Application No. 221091/85), pTerm2 (U.S. Pat. Nos. 4,686,191, 4,939,094 and 5,160,735), pSupex, pUB110, pTP5, pC194 and pEG400 (J. Racteriol., 172:2392 (1990)), PGEX (manufactured by Pharmacia), pET system (manufactured by Novagen), and the like.
[0390] Any promoter can be used so long as it can function in the host cell. Examples include promoters derived from Escherichia coli, phage and the like, such as trp promoter (Ptrp), iac promoter, PL promoter, PR promoter, T7 promoter and the like. Also, artificially designed and modified promoters, such as a promoter in which two Ptrp are linked in series (Ptrp×2), tac promoter, lacT7 promoter letI promoter and the like, can be used.
[0391] It is preferred to use a plasmid in which the space between Shine-Dalgarno sequence which is the ribosome binding sequence and the initiation codon is adjusted to an appropriate distance (for example, 6 to 18 nucleotides).
[0392] The transcription termination sequence is not always necessary for the expression of the DNA of the present invention. However, it is preferred to arrange the transcription terminating sequence at just downstream of the structural gene.
[0393] One of ordinary skill in the art will appreciate that the codons of the above-described elements may be optimized, in a known manner, depending on the host cells and environmental conditions utilized.
[0394] Examples of the host cell include microorganisms belonging to the genus Escherichia, the genus Serratia, the genus Bacillus, the genus Brevibacterium, the genus Corynebacterium, the genus Microbacterinum, the genus Pseudomonas, and the like. Specific examples include Escherichia coli XL1-Blue, Escherichia coli XL2-Blue, Escherichia coli DH1, Escherichia coli MC1000, Escherichia coli KY3276, Escherichia coli W1485, Escherichia coli JM109, Escherichia coli HB101, Escherichia coli No. 49, Escherichia coli W3110, Escherichia coli NY49, Escherichia coli GI698, Escherichia coli TB1, Serratia ficaria, Serratia fonticola, Serratia liquefaciens, Serratia marcescens, Bacillus subtilis, Bacillus amyloliquefaciens, Corynebacterium ammoniagenes, Brevibacterium immariophilum ATCC 14068, Brevibacterium saccharolyticum ATCC 14066, Corynebacterim glutamicum ATCC 13032, Corynebacterium glutamicum ATCC 13869, Corynebacterium glutamicum ATCC 14067 (prior genus and species:Brevibacterium flavum), Corynebacterium glutamicum ATCC 13869 (prior genus and species:Brevibacterium lactofezmentum, or Corynebacterium lactofermentum) Corynebacterium acetoacidophilum ATCC 13870, Corynebacterium thermoaminogenes FERM 9244, Microbacterinum ammoniaphilum ATCC 15354, Pseudomonas putida, Pseudomonas sp. D-0110, and the like.
[0395] When Corynebacterium glutamicum or an analogous microorganism is used as a host, an EMF necessary for expressing the polypeptide is not always contained in the vector so long as the polynucleotide of the present invention contains an ENF When the EMF is not contained in the polynucleotide, it is necessary to prepare the EMF separately and ligate it so as to be in operable combination. Also, when a higher expression amount or specific expression regulation is necessary, it is necessary to ligate the EMF corresponding thereto so as to put the EMF in operable combination with the polynucleotide Examples of using an externally ligated EMF are disclosed in Microbiology, 142:1297-1309 (1996).
[0396] With regard to the method for the introduction of the recombinant vector, the method for introducing DNA into the above-described host cells, such as a method in which a calcium ion is used (Proc. Natl. Acad. Sci. USA, 69:2110 (1972)), a protoplast method (Japanese Published Unexamined Patent Application No. 2483942/88), the methods described in Gene, 17:107 (1982) and Molecular & General Genetics, 168:111 (1979) and the like, can be used.
[0397] When yeast is used as the host cell, examples of the expression vector include pYES2 (manufactured by Invitrogen), YEpl3 (ATCC 37115), YEp24 (ATCC 37051), YCp5O (ATCC 37419), pHSl9, pHS15, and the like.
[0398] Any promoter can be used so long as it can be expressed in yeast. Examples include a promoter of a gene in the glycolytic pathway, such as hexose kinase and the like, PHO5 promoter, PGK promoter, GAP promoter, ADH promoter, gal 1 promoter, gal 10 promoter, a heat shock protein promoter, MF al promoter, CUP 1 promoter, and the like.
[0399] Examples of the host cell include microorganisms belonging to the genus Saccharomyces, the genus Schizosaccharomyces, the genus Kluyveromyces, the genus Trichosporon, the genus Schwanniomyces, the genus Pichia, the genus Candida and the like. Specific examples include Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces lactis, Trichosporon pullulans, Schwanniomyces alluviums, Candida utilis and the like.
[0400] With regard to the method for the introduction of the recombinant vector, any method for introducing DNA into yeast, such as an electroporation method (Methods. Enzymol., 194:182 (1990)), a spheroplast method (Proc. Natl. Acad. Sci. USA, 75:1929 (1978)), a lithium acetate method (J. Bacteriol., 153:163 (1983)), a method described in Proc. Natl. Acad. Sci. USA, 75:1929 (1978) and the like, can be used.
[0401] When animal cells are used as the host cells, examples of the expression vector include pcDNA3.1, pSinRep5 and pCEP4 (manufactured by Invitorogen), pRev-Tre (manufactured by Clontech), pAxCAwt (manufactured by Takara Shuzo), pcDNAI and pcDM8 (manufactured by Funakoshi), pAGE107 (Japanese Published Unexamined Patent Application No. 22979/91; Cytotechnology, 3:133 (1990)), pAS3-3 (Japanese Published Unexamined Patent Application No. 227075/90), pcDM8 (Nature, 329:840 (1987)), pcDNAI/Amp (manufactured by Invitrogen), pREP4 (manufactured by Invitrogen), pAGE103 (J. Biochem., 101:1307 (1987)), pAGE210, and the like.
[0402] Any promoter can be used so long as it can function in animal cells. Examples include a promoter of IE (immediate early) gene of cytomegalovirus (CMV), an early promoter of SV40, a promoter of retrovirus, a metallothionein promoter, a heat shock promoter, SRa promoter, and the like. Also, the enhancer of the IE gene of human CMV can be used together with the promoter.
[0403] Examples of the host cell include human Namalwa cell, monkey COS cell, Chinese hamster CHO cell, HST5637 (Japanese Published Unexamined Patent Application No. 299/88), and the like.
[0404] The method for introduction of the recombinant vector into animal cells is not particularly limited, so long as it is the general method for introducing DNA into animal cells, such as an electroporation method (Cytotechnology, 3:133 (1990)), a calcium phosphate method (Japanese Published Unexamined Patent Application No. 227075/90), a lipofection method (Proc. Natl. Acad. Sci. USA, 84, 7413 (1987)), the method described in Virology, 52:456 (1973), and the like.
[0405] When insect cells are used as the host cells, the polypeptide can be expressed, for example, by the method described in Bacurovirus Expression Vectors, A Laboratory Manual, W. H. Freeman and Company, New York (1992), Bio/Technology, 6:47 (1988), or the like.
[0406] Specifically, a recombinant gene transfer vector and bacurovirus are simultaneously inserted into insect cells to obtain a recombinant virus in an insect cell culture supernatant, and then the insect cells are infected with the resulting recombinant virus to express the polypeptide.
[0407] Examples of the gene introducing vector used in the method include pBlueBac4.5, pVL1392, pVL1393 and pBlueBacIII (manufactured by Invitrogen), and the like.
[0408] Examples of the bacurovirus include Autographa californica nuclear polyhedrosis virus with which insects of the family Barathra are infected, and the like.
[0409] Examples of the insect cells include Spodoptera frugiperda oocytes Sf9 and Sf21 (Bacurovirus Expression Vectors, A Laboratory Manual, W. H. Freeman and Company, New York (1992)), Trichoplusia ni oocyte High 5 (manufactured by Invitrogen) and the like.
[0410] The method for simultaneously incorporating the above-described recombinant gene transfer vector and the above-described bacurovirus for the preparation of the recombinant virus include calcium phosphate method (Japanese Published Unexamined Patent Application No. 227075/90), lipofection method (Proc. Natl. Acad. Sci. USA, 84:7413 (1987)) and the like.
[0411] When plant cells are used as the host cells, examples of expression vector include a Ti plasmid, a tobacco mosaic virus vector, and the like.
[0412] Any promoter can be used so long as it can be expressed in plant cells. Examples include 35S promoter of cauliflower mosaic virus (CaMV), rice actin 1 promoter, and the like.
[0413] Examples of the host cells include plant cells and the like, such as tobacco, potato, tomato, carrot, soybean, rape, alfalfa, rice, wheat, barley, and the like.
[0414] The method for introducing the recombinant vector is not particularly limited, so long as it is the general method for introducing DNA into plant cells, such as the Agrobacterium method (Japanese Published Unexamined Patent Application No. 140885/84, Japanese Published Unexamined Patent Application No. 70080/85, WO 94/00977), the electroporation method (Japanese Published Unexamined Patent Application No. 251887/85), the particle gun method (Japanese Patents 2606856 and 2517813), and the like.
[0415] The transformant of the present invention includes a transformant containing the polypeptide of the present invention per se rather than as a recombinant vector, that is, a transformant containing the polypeptide of the present invention which is integrated into a chromosome of the host, in addition to the transformant containing the above recombinant vector.
[0416] When expressed in yeasts, animal cells, insect cells or plant cells, a glycopolypeptide or glycosylated polypeptide can be obtained.
[0417] The polypeptide can be produced by culturing the thus obtained transformant of the present invention in a culture medium to produce and accumulate the polypeptide of the present invention or any polypeptide expressed under the control of an EMF of the present invention, and recovering the polypeptide from the culture.
[0418] Culturing of the transformant of the present invention in a culture medium is carried out according to the conventional method as used in culturing of the host.
[0419] When the transformant of the present invention is obtained using a prokaryote, such as Escherichia coli or the like, or a eukaryote, such as yeast or the like, as the host, the transformant is cultured.
[0420] Any of a natural medium and a synthetic medium can be used, so long as it contains a carbon source, a nitrogen source, an inorganic salt and the like which can be assimilated by the transformant and can perform culturing of the transformant efficiently.
[0421] Examples of the carbon source include those which can be assimilated by the transformant, such as carbohydrates (for example, glucose, fructose, sucrose, molasses containing them, starch, starch hydrolysate, and the like), organic acids (for example, acetic acid, propionic acid, and the like), and alcohols (for example, ethanol, propanol, and the like).
[0422] Examples of the nitrogen source include ammonia, various ammonium salts of inorganic acids or organic acids (for example, ammonium chloride, ammonium sulfate, ammonium acetate, ammonium phosphate, and the like), other nitrogen-containing compounds, peptone, meat extract, yeast extract, corn steep liquor, casein hydrolysate, soybean meal and soybean meal hydrolysate, various fermented cells and hydrolysates thereof, and the like.
[0423] Examples of inorganic salt include potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium phosphate, magnesium sulfate, sodium chloride, ferrous sulfate, manganese sulfate, copper sulfate, calcium carbonate, and the like.
[0424] The culturing is carried out under aerobic conditions by shaking culture, submerged-aeration stirring culture or the like. The culturing temperature is preferably from 15 to 40° C., and the culturing time is generally from 16 hours to 7 days. The pH of the medium is preferably maintained at 3.0 to 9.0 during the culturing. The pH can be adjusted using an inorganic or organic acid, an alkali solution, urea, calcium carbonate, ammonia, or the like.
[0425] Also, antibiotics, such as ampicillin, tetracycline, and the like, can be added to the medium during the culturing, if necessary.
[0426] When a microorganism transformed with a recombinant vector containing an inducible promoter is cultured, an inducer can be added to the medium, if necessary.
[0427] For example, isopropyl-&bgr;-D-thiogalactopyranoside (IPTG) or the like can be added to the medium when a microorganism transformed with a recombinant vector containing lac promoter is cultured, or indoleacrylic acid (IAA) or the like can by added thereto when a microorganism transformed with an expression vector containing trp promoter is cultured.
[0428] Examples of the medium used in culturing a transformant obtained using animal cells as the host cells include RPMI 1640 medium (The Journal of the American Medical Association, 199:519 (1967)), Eagle's MEM medium (Science, 122:501 (1952)), Dulbecco's modified MEM medium (Virology, 8, 396 (1959)), 199 Medium (Proceeding of the Society for the Biological Medicine, 73:1 (1950)), the above-described media to which fetal calf serum has been added, and the like.
[0429] The culturing is carried out generally at a pH of 6 to 8 and a temperature of 30 to 40° C. in the presence of 5% CO2 for 1 to 7 days.
[0430] Also, if necessary, antibiotics, such as kanamycin, penicillin, and the like, can be added to the medium during the culturing.
[0431] Examples of the medium used in culturing a transformant obtained using insect cells as the host cells include TNM-FH medium (manufactured by Pharmingen), Sf-900 II SFM (manufactured by Life Technologies), ExCell 400 and ExCell 405 (manufactured by JRH Biosciences), Grace's Insect Medium (Nature, 195:788 (1962)), and the like.
[0432] The culturing is carried out generally at a pH of 6 to 7 and a temperature of 25 to 30° C. for 1 to 5 days.
[0433] Additionally, antibiotics, such as gentamicin and the like, can be added to the medium during the culturing, if necessary.
[0434] A transformant obtained by using a plant cell as the host cell can be used as the cell or after differentiating to a plant cell or organ. Examples of the medium used in the culturing of the transformant include Murashige and Skoog (MS) medium, White medium, media to which a plant hormone, such as auxin, cytokinine, or the like has been added, and the like.
[0435] The culturing is carried out generally at a pH of 5 to 9 and a temperature of 20 to 40° C. for 3 to 60 days.
[0436] Also, antibiotics, such as kanamycin, hygromycin and the like, can be added to the medium during the culturing, if necessary.
[0437] As described above, the polypeptide can be produced by culturing a transformant derived from a microorganism, animal cell or plant cell containing a recombinant vector to which a DNA encoding the polypeptide of the present invention has been inserted according to the general culturing method to produce and accumulate the polypeptide, and recovering the polypeptide from the culture.
[0438] The process of gene expression may include secretion of the encoded protein production or fusion protein expression and the like in accordance with the methods described in Molecular Cloning, 2nd ed., in addition to direct expression.
[0439] The method for producing the polypeptide of the present invention includes a method of intracellular expression in a host cell, a method of extracellular secretion from a host cell, or a method of production on a host cell membrane outer envelope. The method can be selected by changing the host cell employed or the structure of the polypeptide produced.
[0440] When the polypeptide of the present invention is produced in a host cell or on a host cell membrane outer envelope, the polypeptide can be positively secreted extracellularly according to, for example, the method of Paulson et al. (J. Biol. Chem., 264:17619 (1989)), the method of Lowe et al. (Proc. Natl. Acad. Sci. USA, 86:8227 (1989); Genes Develop., 4:1288 (1990)), and/or the methods described in Japanese Published Unexamined Patent Application No. 336963/93, Wo 94/23021, and the like.
[0441] Specifically, the polypeptide of the present invention can be positively secreted extracellularly by expressing it in the form that a signal peptide has been added to the foreground of a polypeptide containing an active site of the polypeptide of the present invention according to the recombinant DNA technique.
[0442] Furthermore, the amount produced can be increased using a gene amplification system, such as by use of a dihydrofolate reductase gene or the like according to the method described in Japanese Published Unexamined Patent Application No. 227075/90.
[0443] Moreover, the polypeptide of the present invention can be produced by a transgenic animal individual (transgenic nonhuman animal) or plant individual (transgenic plant).
[0444] When the transformant is the animal individual or plant individual, the polypeptide of the present invention can be produced by breeding or cultivating it so as to produce and accumulate the polypeptide, and recovering the polypeptide from the animal individual or plant individual.
[0445] Examples of the method for producing the polypeptide of the present invention using the animal individual include a method for producing the polypeptide of the present invention in an animal developed by inserting a gene according to methods known to those of ordinary skill in the art (American Journal of Clinical Nutrition, 63:639S (1996), American Journal of Clinical Nutrition, 63:627S (1996), Bio/Technology, 9:830 (1991)).
[0446] In the animal individual, the polypeptide can be produced by breeding a transgenic nonhuman animal to which the DNA encoding the polypeptide of the present invention has been inserted to produce and accumulate the polypeptide in the animal, and recovering the polypeptide from the animal. Examples of the production and accumulation place in the animal include milk (Japanese Published Unexamined Patent Application No. 309192/88), egg and the like of the animal. Any promoter can be used, so long as it can be expressed in the animal. Suitable examples include an &agr;-casein promoter, a &bgr;-casein promoter, a &bgr;-lactoglobulin promoter, a whey acidic protein promoter, and the like, which are specific for mammary glandular cells.
[0447] Examples of the method for producing the polypeptide of the present invention using the plant individual include a method for producing the polypeptide of the present invention by cultivating a transgenic plant to which the DNA encoding the protein of the present invention by a known method (Tissue Culture, 20 (1994), Tissue Culture, 21 (1994), Trends in Biotechnology, 15:45 (1997)) to produce and accumulate the polypeptide in the plant, and recovering the polypeptide from the plant.
[0448] The polypeptide according to the present invention can also be obtained by translation in vitro.
[0449] The polypeptide of the present invention can be produced by a translation system in vitro. There are, for example, two in vitro translation methods which may be used, namely, a method using RNA as a template and another method using DNA as a template. The template RNA includes the whole RNA, mRNA, an in vitro transcription product, and the like. The template DNA includes a plasmid containing a transcriptional promoter and a target gene integrated therein and downstream of the initiation site, a PCR/RT-PCR product and the like. To select the most suitable system for the in vitro translation, the origin of the gene encoding the protein to be synthesized (prokaryotic cell/eucaryotic cell), the type of the template (DNA/RNA), the purpose of using the synthesized protein and the like should be considered. In vitro translation kits having various characteristics are commercially available from many companies (Boehringer Mannheim, Promega, Stratagene, or the like), and every kit can be used in producing the polypeptide according to the present invention.
[0450] Transcription/translation of a DNA nucleotide sequence cloned into a plasmid containing a T7 promoter can be carried out using an in vitro transcription/translation system E. coli T7 S30 Extract System for Circular DNA (manufactured by Promega, catalogue No. L1130). Also, transcription/translation using, as a template, a linear prokaryotic DNA of a supercoil non-sensitive promoter, such as lacUV5, tac, &lgr;PL(con), &lgr;PL, or the like, can be carried out using an in vitro transcription/translation system E. coli S30 Extract System for Linear Templates (manufactured by Promega, catalogue No. L1030). Examples of the linear prokaryotic DNA used as a template include a DNA fragment, a PCR-amplified DNA product, a duplicated oligonucleotide ligation, an in vitro transcriptional RNA, a prokaryotic RNA, and the like.
[0451] In addition to the production of the polypeptide according to the present invention, synthesis of a radioactive labeled protein, confirmation of the expression capability of a cloned gene, analysis of the function of transcriptional reaction or translation reaction, and the like can be carried out using this system.
[0452] The polypeptide produced by the transformant of the present invention can be isolated and purified using the general method for isolating and purifying an enzyme. For example, when the polypeptide of the present invention is expressed as a soluble product in the host cells, the cells are collected by centrifugation after cultivation, suspended in an aqueous buffer, and disrupted using an ultrasonicator, a French press, a Manton Gaulin homogenizer, a Dynomill, or the like to obtain a cell-free extract. From the supernatant obtained by centrifuging the cell-free extract, a purified product can be obtained by the general method used for isolating and purifying an enzyme, for example, solvent extraction, salting out using ammonium sulfate or the like, desalting, precipitation using an organic solvent, anion exchange chromatography using a resin, such as diethylaminoethyl (DEAE)-Sepharose, DIAION HPA-75 (manufactured by Mitsubishi Chemical) or the like, cation exchange chromatography using a resin, such as S-Sepharose FF (manufactured by Pharmacia) or the like, hydrophobic chromatography using a resin, such as butyl sepharose, phenyl sepharose or the like, gel filtration using a molecular sieve, affinity chromatography, chromatofocusing, or electrophoresis, such as isoelectronic focusing or the like, alone or in combination thereof.
[0453] When the polypeptide is expressed as an insoluble product in the host cells, the cells are collected in the same manner, disrupted and centrifuged to recover the insoluble product of the polypeptide as the precipitate fraction. Next, the insoluble product of the polypeptide is solubilized with a protein denaturing agent. The solubilized solution is diluted or dialyzed to lower the concentration of the protein denaturing agent in the solution. Thus, the normal configuration of the polypeptide is reconstituted. After the procedure, a purified product of the polypeptide can be obtained by a purification/isolation method similar to the above.
[0454] When the polypeptide of the present invention or its derivative (for example, a polypeptide formed by adding a sugar chain thereto) is secreted out of cells, the polypeptide or its derivative can be collected in the culture supernatant. Namely, the culture supernatant is obtained by treating the culture medium in a treatment similar to the above (for example, centrifugation). Then, a purified product can be obtained from the culture medium using a purification/isolation method similar to the above.
[0455] The polypeptide obtained by the above method is within the scope of the polypeptide of the present invention, and examples include a polypeptide encoded by a polynucleotide comprising the nucleotide sequence selected from SEQ ID NOS:2 to 3431, and a polypeptide comprising an amino acid sequence represented by any one of SEQ ID NOS:3502 to 6931.
[0456] Furthermore, a polypeptide comprising an amino acid sequence in which at least one amino acids is deleted, replaced, inserted or added in the amino acid sequence of the polypeptide and having substantially the same activity as that of the polypeptide is included in the scope of the present invention. The term “substantially the same activity as that of the polypeptide” means the same activity represented by the inherent function, enzyme activity or the like possessed by the polypeptide which has not been deleted, replaced, inserted or added. The polypeptide can be obtained using a method for introducing part-specific mutation(s) described in, for example, Molecular Cloning, 2nd ed., Current Protocols in Molecular Biology, Nuc. Acids. Res., 10:6487 (1982), Proc. Natl. Acad. Sci. USA, 79:6409 (1982), Gene, 34:315 (1985), Nuc. Acids. Res., 13:4431 (1985), Proc. Natl. Acad. Sci. USA, 82:488 (1985) and the like. For example, the polypeptide can be obtained by introducing mutation(s) to DNA encoding a polypeptide having the amino acid sequence represented by any one of SEQ ID NOS:3502 to 6931. The number of the amino acids which are deleted, replaced, inserted or added is not particularly limited; however, it is usually 1 to the order of tens, preferably 1 to 20, more preferably 1 to 10, and most preferably 1 to 5, amino acids.
[0457] The at least one amino acid deletion, replacement, insertion or addition in the amino acid sequence of the polypeptide of the present invention is used herein to refer to that at least one amino acid is deleted, replaced, inserted or added to at one or plural positions in the amino acid sequence. The deletion, replacement, insertion or addition may be caused in the same amino acid sequence simultaneously. Also, the amino acid residue replaced, inserted or added can be natural or non-natural. Examples of the natural amino acid residue include L-alanine, L-asparagine, L-asparatic acid, L-glutamine, L-glutamic acid, glycine, L-histidine, L-isoleucine, L-leonine, L-lysine, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, L-cysteine, and the like.
[0458] Herein, examples of amino acid residues which are replaced with each other are shown below. The amino acid residues in the samie group can be replaced with each other.
[0459] Group A:
[0460] leonine, isoleucine, norleucine, valine, norvaline, alanine, 2-aminobutanoic acid, methionine, 0-methylserine, t-butylglycine, t-butylalanine, cyclohexylalanine;
[0461] Group B:
[0462] asparatic acid, glutamic acid, isoasparatic acid, isoglutamic acid, 2-aminoadipic acid, 2-aminosuberic acid;
[0463] Group C:
[0464] asparagine, glutamine;
[0465] Group D:
[0466] lysine, arginine, ornithine, 2,4-diaminobutanoic acid, 2,3-diaminopropionic acid;
[0467] Group E:
[0468] proline, 3-hydroxyproline, 4-hydroxyproline;
[0469] Group F:
[0470] serine, threonine, homoserine;
[0471] Group G:
[0472] phenylalanine, tyrosine.
[0473] Also, in order that the resulting mutant polypeptide has substantially the same activity as that of the polypeptide which has not been mutated, it is preferred that the mutant polypeptide has a homology of 60% or more, preferably 80% or more, and particularly preferably 95% or more, with the polypeptide which has not been mutated, when calculated, for example, using default (initial setting) parameters by a homology searching software, such as BLAST, FASTA, or the like.
[0474] Also, the polypeptide of the present invention can be produced by a chemical synthesis method, such as Fmoc (fluorenylmethyloxycarbonyl) method, tboc (t-butyloxycarbonyl) method, or the like. It can also be synthesized using a peptide synthesizer manufactured by Advanced ChemTech, Perkin-Elmer, Pharmacia, Protein Technology Instrument, Synthecell-Vega, PerSeptive, Shimadzu Corporation, or the like.
[0475] The transformant of the present invention can be used for objects other than the production of the polypeptide of the present invention.
[0476] Specifically, at least one component selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof can be produced by culturing the transformant containing the polynucleotide or recombinant vector of the present invention in a medium to produce and accumulate at least one component selected from amino acids, nucleic acids, vitamins, saccharides, organic acids, and analogues thereof, and recovering the same from the medium.
[0477] The biosynthesis pathways, decomposition pathways and regulatory mechanisms of physiologically active substances such as amino acids, nucleic acids, vitamins, saccharides, organic acids and analogues thereof differ from organism to organism. The productivity of such a physiologically active substance can be improved using these differences, specifically by introducing a heterogeneous gene relating to the biosynthesis thereof. For example, the content of lysine, which is one of the essential amino acids, in a plant seed was improved by introducing a synthase gene derived from a bacterium (WO 93/19190). Also, arginine is excessively produced in a culture by introducing an arginine synthase gene derived from Escherichia coli (Japanese Examined Patent Publication 23750/93).
[0478] To produce such a physiologically active substance, the transformant according to the present invention can be cultured by the same method as employed in culturing the transformant for producing the polypeptide of the present invention as described above. Also, the physiologically active substance can be recovered from the culture medium in combination with, for example, the ion exchange resin method, the precipitation method and other known methods.
[0479] Examples of methods known to one of ordinary skill in the art include electroporation, calcium transfection, the protoplast method, the method using a phage, and the like, when the host is a bacterium; and microinjection, calcium phosphate transfection, the positively charged lipid-mediated method and the method using a virus, and the like, when the host is a eukaryote (Molecular Cloning, 2nd ed.; Spector et al., Cells/a laboratory manual, Cold Spring Harbour Laboratory Press, 1998)). Examples of the host include prokaryotes, lower eukaryotes (for example, yeasts), higher eukaryotes (for example, mammals), and cells isolated therefrom. As the state of a recombinant polynucleotide fragment present in the host cells, it can be integrated into the chromosome of the host. Alternatively, it can be integrated into a factor (for example, a plasmid) having an independent replication unit outside the chromosome. These transformants are usable in producing the polypeptides of the present invention encoded by the ORF of the genome of Corynebacterium glutamicum, the polynucleotides of the present invention and fragments thereof. Alternatively, they can be used in producing arbitrary polypeptides under the regulation by an EMF of the present invention.
[0480] 11. Preparation of antibody recognizing the polypeptide of the present invention
[0481] An antibody which recognizes the polypeptide of the present invention, such as a polyclonal antibody, a monoclonal antibody, or the like, can be produced using, as an antigen, a purified product of the polypeptide of the present invention or a partial fragment polypeptide of the polypeptide or a peptide having a partial amino acid sequence of the polypeptide of the present invention.
[0482] (1) Production of polyclonal antibody
[0483] A polyclonal antibody can be produced using, as an antigen, a purified product of the polypeptide of the present invention, a partial fragment polypeptide of the polypeptide, or a peptide having a partial amino acid sequence of the polypeptide of the present invention, and immunizing an animal with the same.
[0484] Examples of the animal to be immunized include rabbits, goats, rats, mice, hamsters, chickens and the like.
[0485] A dosage of the antigen is preferably 50 to 100 &mgr;g per animal.
[0486] When the peptide is used as the antigen, it is preferably a peptide covalently bonded to a carrier protein, such as keyhole limpet haemocyanin, bovine thyroglobulin, or the like. The peptide used as the antigen can be synthesized by a peptide synthesizer.
[0487] The administration of the antigen is, for example, carried out 3 to 10 times at the intervals of 1 or 2 weeks after the first administration. On the 3rd to 7th day after each administration, a blood sample is collected from the venous plexus of the eyeground, and it is confirmed that the serum reacts with the antigen by the enzyme immunoassay (Enzyme-linked Ixmunosorbent Assay (ELISA), Igaku Shoin (1976); Antibodies—A Laboratory Manual, Cold Spring Harbor Laboratory (1988)) or the like.
[0488] Serum is obtained from the immunized non-human mammal with a sufficient antibody titer against the antigen used for the immunization, and the serum is isolated and purified to obtain a polyclonal antibody.
[0489] Examples of the method for the isolation and purification include centrifugation, salting out by 40-50% saturated ammonium sulfate, caprylic acid precipitation (Antibodies, A Laboratory manual, Cold Spring Harbor Laboratory (1988)), or chromatography using a DEAE-Sepharose column, an anion exchange column, a protein A- or G-column, a gel filtration column, and the like, alone or in combination thereof, by methods known to those of ordinary skill in the art.
[0490] (2) Production of monoclonal antibody
[0491] (a) Preparation of antibody-producing cell
[0492] A rat having a serum showing an enough antibody titer against a partial fragment polypeptide of the polypeptide of the present invention used for immunization is used as a supply source of an antibody-producing cell.
[0493] On the 3rd to 7th day after the antigen substance is finally administered the rat showing the antibody titer, the spleen is excised.
[0494] The spleen is cut to pieces in MEM medium (manufactured by Nissui Pharmaceutical), loosened using a pair of forceps, followed by centrifugation at 1,200 rpm for 5 minutes, and the resulting supernatant is discarded.
[0495] The spleen in the precipitated fraction is treated with a Tris-ammonium chloride buffer (pH 7.65) for 1 to 2 minutes to eliminate erythrocytes and washed three times with MEM medium, and the resulting spleen cells are used as antibody-producing cells.
[0496] (b) Preparation of myeloma cells
[0497] As myeloma cells, an established cell line obtained from mouse or rat is used. Examples of useful cell lines include those derived from a mouse, such as P3-X63Ag8-Ul (hereinafter referred to as “P3-U1”) (Curr. Topics in Microbiol. Immunol., 81:1 (1978); Europ. J. Immunol., 6:511 (1976)); SP2/O-Ag14 (SP-2) (Nature, 276:269 (1978)):P3-X63-Ag8653 (653) (J. Immunol., 123:1548 (1979)); P3-X63-Ag8 (X63) cell line (Nature, 256:495 (1975)), and the like, which are 8-azaguanine-resistant mouse (BALB/c) myeloma cell lines. These cell lines are subcultured in 8-azaguanine medium (medium in which, to a medium obtained by adding 1.5 mmol/l glutamine, 5×10−5 mol/l 2-mercaptoethanol, 10 &mgr;g/ml gentamicin and 10% fetal calf serum (FCS) (manufactured by CSL) to RPMI-1640 medium (hereinafter referred to as the “normal medium”), 8-azaguanine is further added at 15 &mgr;g/ml) and cultured in the normal medium 3 or 4 days before cell fusion, and 2×107 or more of the cells are used for the fusion.
[0498] (c) Production of hybridoma
[0499] The antibody-producing cells obtained in (a) and the myeloma cells obtained in (b) are washed with MEM medium or PBS (disodium hydrogen phosphate:1.83 g, sodium dihydrogen phosphate:0.21 g, sodium chloride:7.65 g, distilled water:1 liter, pH:7.2) and mixed to give a ratio of antibody-producing cells:myeloma cells=5:1 to 10:1, followed by centrifugation at 1,200 rpm for 5 minutes, and the supernatant is discarded.
[0500] The cells in the resulting precipitated fraction were thoroughly loosened, 0.2 to 1 ml of a mixed solution of 2 g of polyethylene glycol-1000 (PEG-1000), 2 ml of MEM medium and 0.7 ml of dimethylsulfoxide (DMSO) per 108 antibody-producing cells is added to the cells under stirring at 37° C., and then 1 to 2 ml of MEM medium is further added thereto several times at 1 to 2 minute intervals.
[0501] After the addition, MEM medium is added to give a total amount of 50 ml. The resulting prepared solution is centrifuged at 900 rpm for 5 minutes, and then the supernatant is discarded. The cells in the resulting precipitated fraction were gently loosened and then gently suspended in 100 ml of HAT medium (the normal medium to which 10−4 mol/l hypoxanthine, 1.5×10−5 mol/l thymidine and 4×10−7 mol/l aminopterin have been added) by repeated drawing up into and discharging from a measuring pipette.
[0502] The suspension is poured into a 96 well culture plate at 100 fil/well and cultured at 37° C. for 7 to 14 days in a 5% CO2 incubator.
[0503] After culturing, a part of the culture supernatant is recovered, and a hybridoma which specifically reacts with a partial fragment polypeptide of the polypeptide of the present invention is selected according to the enzyme immunoassay described in Antibodies, A Laboratory manual, Cold Spring Harbor Laboratory, Chapter 14 (1998) and the like.
[0504] A specific example of the enzyme immunoassay is described below.
[0505] The partial fragment polypeptide of the polypeptide of the present invention used as the antigen in the immunization is spread on a suitable plate, is allowed to react with a hybridoma culturing supernatant or a purified antibody obtained in (d) described below as a first antibody, and is further allowed to react with an anti-rat or anti-mouse immunoglobulin antibody labeled with an enzyme, a chemical luminous substance, a radioactive substance or the like as a second antibody for reaction suitable for the labeled substance. A hybridoma which specifically reacts with the polypeptide of the present invention is selected as a hybridoma capable of producing a monoclonal antibody of the present invention.
[0506] Cloning is repeated using the hybridoma twice by limiting dilution analysis (HT medium (a medium in which aminopterin has been removed from HAT medium) is firstly used, and the normal medium is secondly used), and a hybridoma which is stable and contains a sufficient amount of antibody titer is selected as a hybridoma capable of producing a monoclonal antibody of the present invention.
[0507] (d) Preparation of monoclonal antibody
[0508] The monoclonal antibody-producing hybridoma cells obtained in (c) are injected intraperitoneally into 8- to 10-week-old mice or nude mice treated with pristane (intraperitoneal administration of 0.5 ml of 2,6,10,14-tetramethylpentadecane (pristane), followed by 2 weeks of feeding) at 5×106 to 2×106 cells/animal. The hybridoma causes ascites tumor in 10 to 21 days.
[0509] The ascitic fluid is collected from the mice or nude mice, and centrifuged to remove solid contents at 3000 rpm for 5 minutes.
[0510] A monoclonal antibody can be purified and isolated from the resulting supernatant according to the method similar to that used in the polyclonal antibody.
[0511] The subclass of the antibody can be determined using a mouse monoclonal antibody typing kit or a rat monoclonal antibody typing kit. The polypeptide amount can be determined by the Lowry method or by calculation based on the absorbance at 280 nm.
[0512] The antibody obtained in the above is within the scope of the antibody of the present invention.
[0513] The antibody can be used for the general assay using an antibody, such as a radioactive material labeled immunoassay (RIA), competitive binding assay, an immunotissue chemical staining method (ABC method, CSA method, etc.), immunoprecipitation, Western blotting, ELISA assay, and the like (An introduction to Radioimmunoassay and Related Techniques, Elsevier Science (1986); Techniques in Immunocytochemistry, Academic Press, Vol. 1 (1982), Vol. 2 (1983) & Vol. 3 (1985); Practice and Theory of Enzyme Immunoassays, Elsevier Science (1985); Enzyme-linked Immunosorbent Assay (ELISA), Igaku Shoin (1976); Antibodies—A Laboratory Manual, Cold Spring Harbor laboratory (1988); Monoclonal Antibody Experiment Manual, Kodansha Scientific (1987); Second Series Biochemical Experiment Course, Vol. 5, Immunobiochemistry Research Method, Tokyo Kagaku Dojin (1986)).
[0514] The antibody of the present invention can be used as it is or after being labeled with a label.
[0515] Examples of the label include radioisotope, an affinity label (e.g., biotin, avidin, or the like), an enzyme label (e.g., horseradish peroxidase, alkaline phosphatase, or the like), a fluorescence label (e.g., FITC, rhodamine, or the like), a label using a rhodamine atom, (J. Histochem. Cytochem., 18:315 (1970); Meth. Enzym., 62:308 (1979); Immunol., 109:129 (1972); J. Immunol., Meth., 13:215 (1979)), and the like.
[0516] Expression of the polypeptide of the present invention, fluctuation of the expression, the presence or absence of structural change of the polypeptide, and the presence or absence in an organism other than coryneform bacteria of a polypeptide corresponding to the polypeptide can be analyzed using the antibody or the labeled antibody by the above assay, or a polypeptide array or proteome analysis described below.
[0517] Furthermore, the polypeptide recognized by the antibody can be purified by immunoaffinity chromatography using the antibody of the present invention.
[0518] 12. Production and use of polypeptide array
[0519] (1) Production of polypeptide array
[0520] A polypeptide array can be produced using the polypeptide of the present invention obtained in the above item 10 or the antibody of the present invention obtained in the above item 11.
[0521] The polypeptide array of the present invention includes protein chips, and comprises a solid support and the polypeptide or antibody of the present invention adhered to the surface of the solid support.
[0522] Examples of the solid support include plastic such as polycarbonate or the like; an acrylic resin, such as polyacrylamide or the like; complex carbohydrates, such as agarose, sepharose, or the like; silica; a silica-based material, carbon, a metal, inorganic glass, latex beads, and the like.
[0523] The polypeptides or antibodies according to the present invention can be adhered to the surface of the solid support according to the method described in Biotechniques, 27:1258-61 (1999); Molecular Medicine Today, 5:326-7 (1999); Handbook of Experimental Immunology, 4th edition, Blackwell Scientific Publications, Chapter 10 (1986); Meth. Enzym., 34 (1974); Advances in Experimental Medicine and Biology, 42 (1974); U.S. Pat. Nos. 4,681,870; 4,282,287; 4,762,881, or the like.
[0524] The analysis described herein can be efficiently performed by adhering the polypeptide or antibody of the present invention to the solid support at a high density, though a high fixation density is not always necessary.
[0525] (2) Use of polypeptide array
[0526] A polypeptide or a compound capable of binding to and interacting with the polypeptides of the present invention adhered to the array can be identified using the polypeptide array to which the polypeptides of the present invention have been adhered thereto as described in the above (1).
[0527] Specifically, a polypeptide or a compound capable of binding to and interacting with the polypeptides of the present invention can be identified by subjecting the polypeptides of the present invention to the following steps (i) to (iv):
[0528] (i) preparing a polypeptide array having the polypeptide of the present invention adhered thereto by the method of the above (1);
[0529] (ii) incubating the polypeptide immobilized on the polypeptide array together with at least one of a second polypeptide or compound;
[0530] (iii) detecting any complex formed between the at least one of a second polypeptide or compound and the polypeptide immobilized on the array using, for example, a label bound to the at least one of a second polypeptide or compound, or a secondary label which specifically binds to the complex or to a component of the complex after unbound material has been removed; and
[0531] (iv) analyzing the detection data.
[0532] Specific examples of the polypeptide array to which the polypeptide of the present invention has been adhered include a polypeptide array containing a solid support to which at least one of a polypeptide containing an amino acid sequence selected from SEQ ID NOS:3502 to 7001, a polypeptide containing an amino acid sequence in which at least one amino acids is deleted, replaced, inserted or added in the amino acid sequence of the polypeptide and having substantially the same activity as that of the polypeptide, a polypeptide containing an amino acid sequence having a homology of 60% or more with the amino acid sequences of the polypeptide and having substantially the same activity as that of the polypeptides, a partial fragment polypeptide, and a peptide comprising an amino acid sequence of a part of a polypeptide.
[0533] The amount of production of a polypeptide derived from coryneform bacteria can be analyzed using a polypeptide array to which the antibody of the present invention has been adhered in the above (1).
[0534] Specifically, the expression amount of a gene derived from a mutant of coryneform bacteria can be analyzed by subjecting the gene to the following steps (i) to (iv):
[0535] (i) preparing a polypeptide array by the method of the above (1);
[0536] (ii) incubating the polypeptide array (the first antibody) together with a polypeptide derived from a mutant of coryneform bacteria;
[0537] (iii) detecting the polypeptide bound to the polypeptide immobilized on the array using a labeled second antibody of the present invention; and
[0538] (iv) analyzing the detection data.
[0539] Specific examples of the polypeptide array to which the antibody of the present invention is adhered include a polypeptide array comprising a solid support to which at least one of an antibody which recognizes a polypeptide comprising an amino acid sequence selected from SEQ ID NOS:3502 to 7001, a polypeptide comprising an amino acid sequence in which at least one amino acids is deleted, replaced, inserted or added in the amino acid sequence of the polypeptide and having substantially the same activity as that of the polypeptide, a polypeptide comprising an amino acid sequence having a homology of 60% or more with the amino acid sequences of the polypeptide and having substantially the same activity as that of the polypeptides, a partial fragment polypeptide, or a peptide comprising an amino acid sequence of a part of a polypeptide.
[0540] A fluctuation in an expression amount of a specific polypeptide can be monitored using a polypeptide obtained in the time course of culture as the polypeptide derived from coryneform bacteria. The culturing conditions can be optimized by analyzing the fluctuation.
[0541] When a polypeptide derived from a mutant of coryneform bacteria is used, a mutated polypeptide can be detected.
[0542] 13. Identification of useful mutation in mutant by proteome analysis
[0543] Usually, the proteome is used herein to refer to a method wherein a polypeptide is separated by two-dimensional electrophoresis and the separated polypeptide is digested with an enzyme, followed by identification of the polypeptide using a mass spectrometer (Ms) and searching a data base.
[0544] The two dimensional electrophoresis means an electrophoretic method which is perforned by combining two electrophoretic procedures having different principles. For example, polypeptides are separated depending on molecular weight in the primary electrophoresis. Next, the gel is rotated by 90° or 180° and the secondary electrophoresis is carried out depending on isoelectric point. Thus, various separation patterns can be achieved (JIS K 3600 2474).
[0545] In searching the data base, the amino acid sequence information of the polypeptides of the present invention and the recording medium of the present invention provide for in the above items 2 and 8 can be used.
[0546] The proteome-analysis of a coryneform bacterium and its mutant makes it possible to identify a polypeptide showing a fluctuation therebetween.
[0547] The proteome analysis of a wild type strain of coryneform bacteria and a production strain showing an improved productivity of a target product makes it possible to efficiently identify a mutation protein which is useful in breeding for improving the productivity of a target product or a protein of which expression amount is fluctuated.
[0548] Specifically, a wild type strain of coryneform bacteria and a lysine-producing strain thereof are each subjected to the proteome analysis. Then, a spot increased in the lysine-producing strain, compared with the wild type strain, is found and a data base is searched so that a polypeptide showing an increase in yield in accordance with an increase in the lysine productivity can be identified. For example, as a result of the proteome analysis on a wild type strain and a lysine-producing strain, the productivity of the catalase having the amino acid sequence represented by SEQ ID NO:3785 is increased in the lysine-producing mutant.
[0549] As a result that a protein having a high expression level is identified by proteome analysis using the nucleotide sequence information and the amino acid sequence information, of the genome of the coryneform bacteria of the present invention, and a recording medium storing the sequences, the nucleotide sequence of the gene encoding this protein and the nucleotide sequence in the upstream thereof can be searched at the same time, and thus, a nucleotide sequence having a high expression promoter can be efficiently selected.
[0550] In the proteome analysis, a spot on the two-dimentional electrophoresis gel showing a fluctuation is sometimes derived from a modified protein. However, the modified protein can be efficiently identified using the recording medium storing the nucleotide sequence information, the amino acid sequence information, of the genome of coryneform bacteria, and the recording medium storing the sequences, according to the present invention.
[0551] Moreover, a useful mutation point in a useful mutant can be easily specified by searching a nucleotide sequence (nucleotide sequence of promoters, ORF, or the like) relating to the thus identified protein using a recording medium storing the nucleotide sequence information and the amino acid sequence information, of the genome of coryneform bacteria of the present invention, and a recording medium storing the sequences and using a primer designed on the basis of the detected nucleotide sequence. As a result that the useful mutation point is specified, an industrially useful mutant having the useful mutation or other useful mutation derived therefrom can be easily bred.
[0552] The present invention will be explained in detail below based on Examples. However, the present invention is not limited thereto.
EXAMPLE 1[0553] Determination of the full nucleotide sequence of genome of Corynebacterium glutamicum
[0554] The full nucleotide sequence of the genome of Corynebacterium glutamicum was determined based on the whole genome shotgun method (Science, 269:496-512 (1995)). In this method, a genome library was prepared and the terminal sequences were determined at random. Subsequently, these sequences were ligated on a computer to cover the full genome. Specifically, the following procedure was carried out.
[0555] (1) Preparation of genome DNA of Corynebacterium glutamicum ATCC 13032
[0556] Corynebacterium glutamicum ATCC 13032 was cultured in BY medium (7 g/l meat extract, 10 g/l peptone, 3 g/l sodium chloride, 5 g/l yeast extract, pH 7.2) containing 1% of glycine at 30° C. overnight and the cells were collected by centrifugation. After washing with STE buffer (10.3% sucrose, 25 mmol/l Tris hydrochloride, 25 nmuol/l EDTA, pH 8.0), the cells were suspended in 10 ml of STE buffer containing 10 mg/ml lysozyme, followed by gently shaking at 37° C. for 1 hour. Then, 2 ml of 10% SDS was added thereto to lyse the cells, and the resultant mixture was maintained at 65° C. for 10 minutes and then cooled to room temperature. Then, 10 ml of Tris-neutralized phenol was added thereto, followed by gently shaking at room temperature for 30 minutes and centrifugation (15,000×g, 20 minutes, 20° C.). The aqueous layer was separated and subjected to extraction with phenol/chloroform and extraction with chloroform (twice) in the same manner. To the aqueous layer, 3 mol/l sodium acetate solution (pH 5.2) and isopropanol were added at 1/10 times volume and twice volume, respectively, followed by gently stirring to precipitate the genome DNA. The genome DNA was dissolved again in 3 ml of TE buffer (10 mmol/l Tris hydrochloride, 1 mmol/l EDTA, pH 8.0) containing 0.02 mg/ml of RNase and maintained at 37° C. for 45 minutes. The extractions with phenol, phenol/chloroform and chloroform were carried out successively in the same manner as the above. The genome DNA was subjected to isopropanol precipitation. The thus formed genome DNA precipitate was washed with 70% ethanol three times, followed by air-drying, and dissolved in 1.25 ml of TE buffer to give a genome DNA solution (concentration:0.1 mg/ml).
[0557] (2) Construction of a shotgun library
[0558] TE buffer was added to 0.01 mg of the thus prepared genome DNA of Corynebacterium glutamicum ATCC 13032 to give a total volume of 0.4 ml, and the mixture was treated with a sonicator (Yamato Powersonic Model 150) at an output of 20 continuously for 5 seconds to obtain fragments of 1 to 10 kb. The genome fragments were blunt-ended using a DNA blunting kit (manufactured by Takara Shuzo) and then fractionated by 6% polyacrylamide gel electrophoresis. Genome fragments of 1 to 2 kb were cut out from the gel, and 0.3 ml MG elution buffer (0.5 mol/l ammonium acetate, 10 mmol/l magnesium acetate, 1 mmol/l EDTA, 0.1% SDS) was added thereto, followed by shaking at 37° C. overnight to elute DNA. The DNA eluate was treated with phenol/chloroform, and then precipitated with ethanol to obtain a genome library insert. The total insert and 500 ng of pUC18 SmaIl/BAP (manufactured by Amersham Pharmacia Biotech) were ligated at 16° C. for 40 hours.
[0559] The ligation product was precipitated with ethanol and dissolved in 0.01 ml of TE buffer. The ligation solution (0.001 ml) was introduced into 0.04 ml of E. coli ELECTRO MAX DH10B (manufactured by Life Technologies) by the electroporation under conditions according to the manufacture's instructions. The mixture was spread on LB plate medium (LB medium (10 g/l bactotrypton, 5 g/l yeast extract, 10 g/l sodium chloride, pH 7.0) containing 1.6% of agar) containing 0.1 mg/ml ampicillin, 0.1 mg/ml X-gal and 1 mmol/l isopropyl-&bgr;-D-thiogalactopyranoside (IPTG) and cultured at 37° C. overnight.
[0560] The transformant obtained from colonies formed on the plate medium was stationarily cultured in a 96-well titer plate having 0. 05 ml of LB medium containing 0.1 mg/ml ampicillin at 37° C. overnight. Then, 0.05 ml of LB medium containing 20% glycerol was added thereto, followed by stirring to obtain a glycerol stock.
[0561] (3) Construction of cosmid library
[0562] About 0.1 mg of the genome DNA of Corynebacterium glutamicum ATCC 13032 was partially digested with Sau3AI (manufactured by Takara Shuzo) and then ultracentrifuged (26,000 rpm, 18 hours, 20° C.) under 10 to 40% sucrose density gradient obtained using 10% and 40% sucrose buffers (1 mol/l NaCl, 20 numol/l Tris hydrochloride, 5 mmol/l EDTA, 10% or 40% sucrose, pH 8.0). After the centrifugation, the solution thus separated was fractionated into tubes at 1 ml in each tube. After confirming the DNA fragment length of each fraction by agarose gel electrophoresis, a fraction containing a large amount of DNA fragment of about 40 kb was precipitated with ethanol.
[0563] The DNA fragment was ligated to the BamHI site of superCosl (manufactured by Stratagene) in accordance with the manufacture's instructions. The ligation product was incorporated into Escherichia coli XL-1-BlueMR strain (manufactured by Stratagene) using Gigapack III Gold Packaging Extract (manufactured by Stratagene) in accordance with the manufacture's instructions. The Escherichia coli was spread on LB plate medium containing 0.1 mg/ml ampicillin and cultured therein at 37° C. overnight to isolate colonies. The resulting colonies were stationarily cultured at 37° C. overnight in a 96-well titer plate containing 0.05 ml of the LB medium containing 0.1 mg/ml ampicillin in each well. L medium containing 20% glycerol (0.05 ml) was added thereto, followed by stirring to obtain a glycerol stock.
[0564] (4) Determination of nucleotide sequence
[0565] (4-1) Preparation of template
[0566] The full nucleotide sequence of Corynebacterium glutamicum ATCC 13032 was determined mainly based on the whole genome shotgun method. The template used in the whole geenome shotgun method was prepared by the PCR method using the library prepared in the above (2).
[0567] Specifically, the clone derived from the whole genome shotgun library was inoculated using a replicator (manufactured by GENETIX) into each well of a 96-well plate containing the LB medium containing 0.1 mg/ml of ampicillin at 0.08 ml per each well and then stationarily cultured at 37° C. overnight.
[0568] Next, the culturing solution was transported using a copy plate (manufactured by Tokken) into a 96-well reaction plate (manufactured by PE Biosystems) containing a PCR reaction solution (TaKaRa Ex Taq (manufactured by Takara Shuzo)) at 0.08 ml per each well. Then, PCR was carried out in accordance with the protocol by Makino et al. (DNA Research, 5:1-9 (1998)) using GeneAmp PCR System 9700 (manufactured by PE Riosystems) to amplify the inserted fragment.
[0569] The excessive primers and nucleotides were eliminated using a kit for purifying a PCR production (manufactured by Amersham Pharmacia Biotech) and the residue was used as the template in the sequencing reaction.
[0570] Some nucleotide sequences were determined using a double-stranded DNA plasmid as a template.
[0571] The double-stranded DNA plasmid as the template was obtained by the following method.
[0572] The clone derived from the whole genome shotgun library was inoculated into a 24- or 96-well plate containing a 2× YT medium (16 g/l bactotrypton, 10 g/l yeast extract, 5 g/l sodium chloride, pH 7 0) containing 0.05 mg/ml ampicillin at 1. 5 ml per each well and then cultured under shaking at 37° C. overnight.
[0573] The double-stranded DNA plasmid was prepared from the culturing solution using an automatic plasmid preparing machine, K BO PI-50 (manufactured by Kurabo Industries) or a multiscreen (manufactured by Millipore) in accordance with the protocol provided by the manufacturer.
[0574] To purify the double-stranded DNA plasmid using the multiscreen, Biomek 2000 -(manufactured by Beckman Coulter) or the like was employed.
[0575] The thus obtained double-stranded DNA plasmid was dissolved in water to give a concentration of about 0.1 mg/ml and used as the template in sequencing.
[0576] (4-2) Sequencing reaction
[0577] To 6 &mgr;l of a solution of ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (manufactured by PE Biosystems), an M13 regular direction primer (M13-21) or an M13 reverse direction primer (M13REV) (DNA Research, 5:1-9 (1998) and the template prepared in the above (4-1) (the PCR product or the plasmid) were added to give 10 &mgr;l of a sequencing reaction solution. The primers and the templates were used in an amount of 1.6 pmol and an amount of 50 to 200 ng, respectively.
[0578] Dye terminator sequencing reaction of 45 cycles was carried out with GeneAmp PCR System 9700 (manufactured by PE Biosystems) using the reaction solution. The cycle parameter was determined in accordance with the manufacturer's instruction accompanying ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit. The sample was purified using MultiScreen HV plate (manufactured by Millipore) according to the manufacture's instructions. The thus purified reaction product was precipitated with ethanol, followed by drying, and then stored in the dark at −30° C.
[0579] The dry reaction product was analyzed by ABI PRISM 377 DNA Sequencer and ABI PRISM 3700 DNA Analyzer (both manufactured by PE Biosystems) each in accordance with the manufacture's instructions.
[0580] The data of about 50,000 sequences in total (i.e., about 42,000 sequences obtained using 377 DNA Sequencer and about 8,000 reactions obtained by 3700 DNA Analyser) were transferred to a server (Alpha Server 4100:manufactured by COMPAQ) and stored. The data of these about 50,000 sequences corresponded to 6 times as much as the genome size.
[0581] (5) Assembly
[0582] All operations were carried out on the basis of UNIX platform. The analytical data were output in Macintosh platform using X Window System. The base call was carried out using phred (The University of Washington). The vector sequence data was deleted using SPS Cross_Match (manufactured by Southwest Parallel Software). The assembly was carried out using SPS phrap (manufactured by Southwest Parallel Software; a high-speed version of phrap (The University of Washington)). The contig obtained by the assembly was analyzed using a graphical editor, consed (The University of Washington). A series of the operations from the base call to the assembly were carried out simultaneously using a script phredPhrap attached to consed.
[0583] (6) Determination of nucleotide sequence in gap part
[0584] Each cosmid in the cosmid library constructed in the above (3) was prepared by a method similar to the preparation of the double-stranded DNA plasmid described in the above (4-1). The nucleotide sequence at the end of the inserted fragment of the cosmid was determined by using ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (manufactured by PE Biosystems) according to the manufacture's instructions.
[0585] About 800 cosmid clones were sequenced at both ends to search a nucleotide sequence in the contig derived from the shotgun sequencing obtained in the above (5) coincident with the sequence. Thus, the linkage between respective cosmid clones and respective contigs were determined and mutual alignment was carried out. Furthermore, the results were compared with the physical map of Corynebacterium glutamicum ATCC 13032 (Mol. Gen. Genet., 252:255-265 (1996) to carrying out mapping between the cosmids and the contigs.
[0586] The sequence in the region which was not covered with the contigs was determined by the following method.
[0587] Clones containing sequences positioned at the ends of contigs were selected. Among these clones, about 1,000 clones wherein only one end of the inserted fragment had been determined were selected and the sequence at the opposite end of the inserted fragment was determined. A shotgun library clone or a cosmid clone containing the sequences at the respective ends of the inserted fragment in two contigs was identified, the full nucleotide sequence of the inserted fragment of this clone was determined, and thus the nucleotide sequence of the gap part was determined. When no shotgun library clone or cosmid clone covering the gap part was available, primers complementary to the end sequences at the two contigs were prepared and the DNA fragment in the gap part was amplified by PCR. Then, sequencing was performed by the primer walking method using the amplified DNA fragment as a template or by the shotgun method in which the sequence of a shotgun clone prepared from the amplified DNA fragment was determined. Thus, the nucleotide sequence of the domain was determined.
[0588] In a region showing a low sequence precision, primers were synthesized using AUTOFINISH function and NAVIGATING function of consed (The University of Washington) and the sequence was determined by the primer walking method to improve the sequence precision. The thus determined full nucleotide sequence of the genome of Corynebacterium glutamicum ATCC 13032 strain is shown in SEQ ID NO:1.
[0589] (7) Identification of ORF and presumption of its function
[0590] ORFs in the nucleotide sequence represented by SEQ ID NO:1 were identified according to the following method. First, the ORF regions were determined using software for identifying ORF, i.e., Glimmer, Generk and GeMark.hmm on UNIX platform according to the respective manual attached to the software.
[0591] Based on the data thus obtained, ORFs in the nucleotide sequence represented by SEQ ID NO:1 were identified.
[0592] The putative function of an ORF was determined by searching the homology of the identified amino acid sequence of the OR against an amino acid database consisting of protein-encoding domains derived from Swiss-Prot, PIR or Genpept database constituted by protein encoding domains derivezd from Gersank database, Frame Search (manufactured by Compugen), or by searching the homology of the identified amino acid sequence of the ORF against an amino acid database consisting of protein-encoding domains derived from Swiss-Prot, PIR or Genpept database constituted by protein encoding domains derived from GenBank database, BLAST The nucleotide sequences of the thus determined ORFs are shown in SEQ ID NOS:2 to 3501, and the amino acid sequences encoded by these ORFs are shown in SEQ ID NOS:3502 to 7001.
[0593] In some cases of the sequence listings in the present invention, nucleotide sequences, such as TTG, TOT, GGT, and the like, other than ATG, are read as an initiating codon encoding Met.
[0594] Also, the preferred nucleotide sequences are SEQ ID NOS:2 to 355 and 357 to 3501, and the preferred amino acid sequences are shown in SEQ ID NOS:3502 to 3855 and 3857 to 7001
[0595] Table 1 shows the registration numbers in the above-described databases of sequences which were judged as having the highest homology with the nucleotide sequences of the ORFs as the results of the homology search in the amino acid sequences using the homology-searching software Frame Search (manufactured by Compugen) names of the genes of these sequences, the functions of the genes, and the matched length, identities and analogies compared with publicly known amino acid-translation sequences Moreover, the corresponding positions were confirmed via the alignment of the nucleotide sequence of an arbitrary ORF with the nucleotide sequence of SEQ ID NO:1. Also, the positions of nucleotide sequences other than the ORFs (for example, ribosomal RNA genes, transfer RN genes, IS sequences, and the like) on the genome were determined.
[0596] FIG. 1 shows the positions of typical genes of the Corynebacterium glutamicum ATCC 13032 on the genome. 1 TABLE 1 SEQ NO. SEQ NO. Identity Similarity Matched (DNA) (a.a.) Initial (nt) Terminal (nt) ORF (bp) db Match Homologous gene (%) (%) length (a.a.) Function 2 3502 1 1572 1572 gsp:R98523 Brevibacterium flavum dnaA 99.8 99.8 524 replication initiation protein DnaA 3 3503 1920 1597 324 4 3504 2292 3473 1182 sp:DP3B_MYCSM Mycobacterium smegmatis dnaN 50.5 81.8 390 DNA polymerase III beta chain 5 3505 3585 4766 1182 sp:RECF_MYCSM Mycobacterium smegmatis recF 53.3 79.9 392 DNA replication protein (recF protein) 6 3506 4766 5299 534 sp:YREG_STRCO Streptomyces coelicolor yreG 35.1 58.1 174 hypothetical protein 7 3507 5354 7486 2133 pir.S44198 Mycobacterium tuberculosis 71.9 88.9 704 DNA topoisomerase (ATP- H37Rv gyrB hydrolyzing) 8 3508 7830 8795 966 9 3509 9466 8798 669 10 3510 9562 10071 510 11 3511 9914 9474 441 12 3512 11177 10107 1071 sp:YV11_MYCTU Mycobacterium tuberculosis 29.4 50.7 422 NAGC/XYLR repressor H37Rv 13 3513 11523 11263 261 14 3514 11768 11523 246 15 3515 11831 14398 2568 sp:GYRA_MYCTU Mycobacterium tuberculosis 70.4 88.1 854 DNA gyrase subunit A H37Rv Rv0006 gyrA 16 3516 14405 14746 342 pir.E70698 Mycobacterium tuberculosis 29.5 69.6 112 hypothetical membrane protein H37Rv Rv0007 17 3517 16243 15209 1035 sp:YEIH_ECOLI Escherichia coli K12 yeiH 33.7 63.5 329 hypothetical protein 18 3518 16314 17207 894 gp:AB042619_1 Hydrogenophilus thermoluteolus 27.6 62.3 268 bacterial regulatory protein, LysR TH-1 cbbR type 19 3519 17251 17670 420 20 3520 18729 17860 870 gp:AF156103_2 Rhodobacter capsulatus ccdA 29.1 57.4 265 cytochrome c biogenesis protein 21 3521 19497 18736 762 pir:A49232 Coxiella burnetii com1 31.6 64.5 155 hypothetical protein 22 3522 19705 20073 369 pir:F70664 Mycobacterium tuberculosis 36.8 70.1 117 repressor H37Rv Rv1846c 23 3523 20073 21065 993 gp:MLCB1788_6 Mycobacterium leprae 24.9 50.8 321 hypothetical membrane protein MLCB1788.18 24 3524 21253 21074 180 pir:I40838 Corynebacterium sp. ATCC 65.4 88.5 26 2,5-diketo-D-gluconic acid reductase 31090 25 3525 21597 22124 528 sp:5NTD_VIBPA Vibrio parahaemolyticus nutA 27.0 56.1 196 5′-nucleotidase precursor 26 3526 22164 23399 1236 gp:AE001909_7 Deinococcus radiodurans 27.0 56.7 270 5′-nucleotidase family protein DR0505 27 3527 23779 23615 165 prf:2513302C Corynebacterium striatum ORF1 52.9 72.6 51 transposase 28 3528 24295 24729 435 prf:2413353A Xanthomonas campestris 51.8 79.9 139 organic hydroperoxide detoxication phaseoli ohr enzyme 29 3529 26297 24885 1413 sp:RECG_THIFE Thiobacillus ferrooxidans recG 32.7 60.8 217 ATP-dependent DNA helicase 30 3530 26338 26775 438 31 3531 28099 26822 1278 sp:AMYH_YEAST Saccharomyces cerevisiae 26.7 54.1 449 glucan 1,4-alpha-glucosidase S288C YIR019C sta1 32 3532 29117 28164 954 gp:ERU52850_1 Erysipelothrix rhusiopathiae 28.9 63.7 311 lipoprotein ewlA 33 3533 29965 29117 849 gp:AF180520_3 Streptococcus pyogenes SF370 34.6 74.1 266 ABC 3 transport family or integral mtsC membrane protein 34 3534 29995 30651 657 sp:FECE_ECOLI Escherichia coli K12 fecE 39.2 70.3 222 iron(III) dicitrate transport ATP- biding protein 35 3535 30697 31677 981 pir:A72417 Thermotoga maritima MSB8 25.8 56.5 283 sugar ABC transporter, periplasmic TM0114 sugar-binding protein 36 3536 31677 32699 1023 prf:1207243B Escherichia coli K12 rbsC 30.5 68.3 312 high affinity ribose transport protein 37 3537 32699 33457 759 sp:RBSA_BACSU Bacillus subtilis 168 rbsA 32.2 76.7 236 ribose transport ATP-binding protein 38 3538 34280 33465 816 pir:151116 Petromyzon marinus 23.6 44.4 347 neurofilament subunit NF-180 39 3539 34339 34899 561 sp:CYPA_MYCTU Mycobacterium leprae H37RV 79.9 89.9 169 peptidyl-prolyl cis-trans isomerase A RV0009 ppiA 40 3540 34982 35668 687 sp:YQGP_BACSU Bacillus subtilis 168 yqgP 29.2 53.1 226 hypothetical membrane protein 41 3541 37221 38198 978 sp:FEPG_ECOLI Escherichia coli K12 fepG 40.4 70.5 332 ferric enterobactin transport system permease protein 42 3542 37242 36247 996 43 3543 38202 38978 777 gp:VCU52150_9 Vibrio cholerae viuC 51.8 81.8 253 ATPase 44 3544 38978 39799 822 sp:VIUB_VIBVU Vibrio vulnificus MO6-24 viuB 26.2 52.7 260 vulnibactin utilization protein 45 3545 40458 40189 270 sp:YO11_MYCTU Mycobacterium tuberculosis 40.0 72.6 95 hypothetical membrane protein H37Rv Rv0011c 46 3546 42513 40576 1938 sp:PKNB_MYCLE Mycobacterium leprae pknB 40.6 68.7 648 serine/threonine protein kinase 47 3547 43919 42513 1407 gp:AF094711_1 Streptomyces coelicolor pksC 31.7 59.1 486 serine/threonine protein kinase 48 3548 45347 43926 1422 gp:AF241575_1 Streptomyces griseus pbpA 33.5 66.7 492 penicillin-binding protein 49 3549 46489 45347 1143 sp:SP5E_BACSU Bacillus subtilis 168 spoVE 31.2 65.6 375 stage V sporulation protein E 50 3550 48021 46669 1353 pir:H70699 Mycobacterium tuberculosis 44.1 70.8 469 phosphoprotein phosphatase H37Rv ppp 51 3551 48485 48024 462 pir:A70700 Mycobacterium tuberculosis 38.7 66.5 155 hypothetical protein H37Rv Rv0019c 52 3552 49368 48505 864 pir:B70700 Mycobacterium tuberculosis 23.6 38.8 526 hypothetical protein H37Rv Rv0020c 53 3553 49601 49455 147 54 3554 50616 49897 720 55 3555 50972 50754 219 56 3556 51436 50966 471 57 3557 53055 54008 954 sp:PH2M_TRICU Trichosporon cutaneum ATCC 29.9 63.3 117 phenol 2-monooxygenase 46490 58 3558 53095 51626 1470 sp:GABD_ECOLI Escherichia coli K12 gabD 46.7 78.2 490 succinate-semialdehyde dehydrogenase (NAD(P)+) 59 3559 54080 55546 1467 sp:YRKH_BACSU Bacillus subtilis yrkH 27.3 57.0 242 hypothetical protein 60 3560 56417 55629 789 sp:Y441_METJA Methanococcus jannaschii 29.0 64.1 262 hypothetical membrane protein MJ0441 61 3561 56676 56386 291 sp:YRKF_BACSU Bacillus subtilis yrkF 40.5 74.3 74 hypothetical protein 62 3562 57270 56680 591 sp:YC61_SYNY3 Synechocystis sp. PCC6803 36.3 70.4 179 hypothetical protein slr1261 63 3563 57478 57651 174 pir:G70988 Mycobacterium tuberculosis 53.2 83.9 62 hypothetical protein H37Rv Rv1766 64 3564 58087 58941 855 65 3565 59091 59930 840 gp:LMFL4768_11 Leishmania major L4768.11 26.8 50.7 310 hypothetical protein 66 3566 59952 60662 711 67 3567 60669 62321 1653 68 3568 63508 62390 1119 pir:F70952 Mycobacterium tuberculosis 29.5 59.5 390 magnesium and cobalt transport H37Rv Rv1239c corA protein 69 3569 64040 63594 447 70 3570 64190 65458 1269 gp:AF179611_12 Zymomonas mobilis ZM4 clcb 30.0 64.8 400 chloride channel protein 71 3571 66197 65508 690 sp:PNUC_SALTY Salmonella typhimurium pnuC 24.1 53.1 241 required for NMN transport 72 3572 66851 67972 1122 sp:PHOL_MYCTU Mycobacterium tuberculosis 29.1 60.0 340 phosphate starvation-induced H37Rv Rv2368C protein-like protein 73 3573 68170 68301 132 74 3574 68634 68251 384 75 3575 69060 69824 765 76 3576 70186 68720 1467 sp:CITM_BACSU Bacillus subtilis citM 42.3 68.8 497 Mg(2+)/citrate complex secondary transporter 77 3577 70506 72158 1653 sp:DPIB_ECOLI Escherichia coli K12 dpiB 27.2 60.6 563 two-component system sensor histidine kinase 78 3578 72043 71474 570 79 3579 72161 72814 654 sp:DPIA_ECOLI Escherichia coli K12 criR 33.2 63.3 229 transcriptional regulator 80 3580 73728 72817 912 gp:AF134895_1 Corynebacterium glutamicum 43.3 73.7 293 D-isomer specific 2-hydroxyacid unkdh dehydrogenase 81 3581 73844 74272 429 gp:SCM2_3 Streptomyces coelicolor A3(2) 38.6 76.4 127 hypothetical protein SCM2.03 82 3582 74490 75491 1002 sp:BIOB_CORGL Corynebacterium glutamicum 99.4 99.7 334 biotin synthase bioB 83 3583 75506 75742 237 pir:H70542 Mycobacterium tuberculosis 72.1 79.1 43 hypothetical protein H37Rv Rv1590 84 3584 75697 76035 339 sp:YK14_YEAST Saccharomyces cerevisiae 34.1 63.5 85 hypothetical protein YKL084w 85 3585 76353 76469 117 86 3586 80753 80613 141 PIR:F81737 Chlamydia muridarum Nigg 71.0 75.0 42 hypothetical protein TC0129 87 3587 81274 81002 273 GSP:Y35814 Chlamydia pneumoniae 61.0 66.0 84 hypothetical protein 88 3588 83568 82120 1449 prf:2512333A Streptomyces virginiae varS 25.6 59.0 507 integral membrane efflux protein 89 3589 84935 83691 1245 gp:D38505_1 Bacillus sp. 97.2 99.8 394 creatinine deaminase 90 3590 85403 85098 306 91 3591 86277 85663 615 92 3592 86318 87241 924 sp:HST2_YEAST Saccharomyces cerevisiae hst2 26.2 50.2 279 SIR2 gene family (silent information regulator) 93 3593 88532 87561 972 prf:2316378A Propionibacterium acnes 30.7 59.0 251 triacylglycerol lipase 94 3594 89444 88545 900 prf.2316378A Propionibacterium acnes 29.4 56.1 262 triacylglycerol lipase 95 3595 89558 90445 888 96 3596 90973 90461 513 gp:AB029154_1 Corynebacterium glutamicum 90.6 94.7 171 transcriptional regulator ureR 97 3597 91174 91473 300 gp:AB029154_2 Corynebacterium glutamicum 100.0 100.0 100 urease gammma subunit or urease ureA structural protein 98 3598 91503 91988 486 gp:CGL251883_2 Corynebacterium glutamicum 100.0 100.0 162 urease beta subunit ATCC 13032 ureB 99 3599 91992 93701 1710 gp:CGL251883_3 Corynebacterium glutamicum 100.0 100.0 570 urease alpha subunit ATCC 13032 ureC 100 3600 93729 94199 471 gp:CGL251883_4 Corynebacterium glutamicum 100.0 100.0 157 urease accessory protein ATCC 13032 ureE 101 3601 94202 94879 678 gp:CGL251883_5 Corynebacterium glutamicum 100.0 100.0 226 urease accessory protein ATCC 13032 ureF 102 3602 94899 95513 615 gp:CGL251883_6 Corynebacterium glutamicum 100.0 100.0 205 urease accessory protein ATCC 13032 ureG 103 3603 95517 96365 849 gp:CGL251883_7 Corynebacterium glutamicum 100.0 100.0 283 urease accessory protein ATCC 13032 ureD 104 3604 97144 96368 777 prf:2318326B Agrobacterium radiobacter echA 21.2 48.4 279 epoxide hydrolase 105 3605 97521 98189 669 106 3606 98470 97319 1152 gp:AF148322_1 Streptomyces viridifaciens vlmF 26.5 59.7 347 valanimycin resistant protein 107 3607 99819 100493 675 108 3608 101582 98808 2775 109 3609 103435 101612 1824 sp:HTPG_ECOLI Escherichia coli K12 htpG 23.8 52.7 668 heat shock protein (hsp90-family) 110 3610 103494 104909 1416 sp:AMN_ECOLI Escherichia coli K12 amn 41.0 68.2 481 AMP nucleosidase 111 3611 105751 105173 579 112 3612 106392 105841 552 pir.E72483 Aeropyrum pernix K1 APE2509 29.6 58.7 196 acetolactate synthase large subunit 113 3613 107289 106630 660 114 3614 107435 110890 3456 sp:PUTA_SALTY Salmonella typhimurium putA 25.8 50.4 1297 proline dehydrogenase/P5C dehydrogenase 115 3615 111161 111274 114 116 3616 111374 112318 945 sp:AAD_PHACH Phanerochaete chrysosporium 30.2 60.7 338 aryl-alcohol dehydrogenase aad (NADP+) 117 3617 112470 114083 1614 sp:YDAH_ECOLI Escherichia coli K12 ydaH 36.5 71.4 513 pump protein (transport) 118 3618 114147 115478 1332 prf:2422424A Enterobacter agglomerans 23.0 49.2 352 indole-3-acetyl-Asp hydrolase 119 3619 115262 114564 699 120 3620 115578 115943 366 sp:YIDH_ECOLI Escherichia coli K12 yidH 35.9 70.8 106 hypothetical membrane protein 121 3621 115949 116263 315 122 3622 118599 116548 2052 123 3623 119589 118810 780 sp:ACCR_AGRTU Agrobacterium tumefaciens 29.5 59.7 258 transcriptional repressor accR 124 3624 120021 120410 390 pir:C70019 Bacillus subtilis yurT 57.9 78.6 126 methylglyoxalase 125 3625 120922 120413 510 sp:YC76_MYCTU Mycobacterium tuberculosis 37.0 64.8 162 hypothetical protein H37Rv Rv1276c 126 3626 122459 120951 1509 prf:2309180A Pseudomonas fluorescens mtlD 43.5 70.4 497 mannitol dehydrogenase 127 3627 123841 122507 1335 prf:2321326A Klebsiella pneumoniae dalT 30.3 68.3 435 D-arabinitol transporter 128 3628 123842 124030 189 129 3629 124130 124966 837 sp:GATR_ECOLI Escherichia coli K12 gatR 27.3 64.6 260 galactitol utilization operon repressor 130 3630 124932 126350 1419 sp:XYLB_STRRU Streptomyces rubiginosus xylB 45.0 68.1 451 xylulose kinase 131 3631 127171 127992 822 132 3632 127189 126353 837 gp:CGPAN_2 Corynebacterium glutamicum 100.0 100.0 279 pantoate-beta-alanine ligase ATCC 13032 panC 133 3633 128004 127192 813 gp:CGPAN_1 Corynebacterium glutamicum 100.0 100.0 271 3-methyl-2-oxobutanoate ATCC 13032 panB hydroxymethyltransferase 134 3634 129049 128099 951 135 3635 130118 129489 630 sp:3MG_ARATH Arabidopsis thaliana mag 42.0 67.6 188 DNA-3-methyladenine glycosylase 136 3636 130145 130798 654 137 3637 131738 130815 924 gp:AB029896_1 Petroleum-degrading bacterium 39.3 69.3 270 esterase HD-1 hde 138 3638 131798 132424 627 139 3639 132424 132981 558 sp:CAH_METTE Methanosarcina thermophila 30.9 53.2 201 carbonate dehydratase 140 3640 134113 132971 1143 sp:XYlR_BACSU Bacillus subtilis W23 xylR 24.1 49.3 357 xylose operon repressor protein 141 3641 135478 134207 1272 gp:LLLPK214_12 Lactococcus lactis mef214 21.1 61.2 418 macrolide efflux protein 142 3642 136321 135518 804 143 3643 136565 136122 444 144 3644 136804 138744 1941 145 3645 138791 140329 1539 146 3646 139861 139226 636 147 3647 140329 141789 1461 pir:I39714 Agrobacterium tumefaciens celA 24.3 51.2 420 cellulose synthase 148 3648 141796 143526 1731 sp:HKR1_YEAST Saccharomyces cerevisiae 25.1 51.8 593 hypothetical membrane protein YDR420W hkr1 149 3649 142455 143075 621 150 3650 143575 144639 1065 151 3651 144725 145480 756 152 3652 146396 145518 879 sp:RARD_PSEAE Pseudomonas aeruginosa rarD 34.7 60.7 303 chloramphenicol sensitive protein 153 3653 146522 147238 717 sp:YADS_ECOLI Escherichia coli Kl2 yadS 30.3 59.1 198 hypothetical membrane protein 154 3654 147238 147570 333 155 3655 148122 149780 1659 156 3656 150930 149794 1137 sp:ABRB_ECOLI Escherichia coli K12 abrB 32.4 62.3 361 transport protein 157 3657 151572 152369 798 sp:YFCA_ECOLI Escherichia coli K12 yfcA 34.7 70.2 248 hypothetical membrane protein 158 3658 151589 150966 624 159 3659 152410 152814 405 160 3660 155613 153226 2388 sp:HRPB_ECOLI Escherichia coli K12 hrpB 33.8 64.3 829 ATP-dependent helicase 161 3661 155853 156167 315 162 3662 156821 156147 675 sp:NODL_RHILV Rhizobium leguminosarum bv. 40.4 66.0 188 nodulation protein viciae plasmid pRL1JI nodL 163 3663 156848 157537 690 sp:ALKB_ECOLI Escherichia coli o373#1 alkB 34.7 60.7 219 DNA repair system specific for alkylated DNA 164 3664 157614 158138 525 sp:3MG1_ECOLI Escherichia coli K12 tag 39.8 65.1 166 DNA-3-methyladenine glycosylase 165 3665 158154 158831 678 sp:RHTC_ECOLI Escherichia coli K12 rhtC 34.1 61.3 217 threonine efflux protein 166 3666 158869 159159 291 sp:YAAA_BACSU Bacillus subtilis yaaA 50.9 72.7 55 hypothetical protein 167 3667 159162 160013 852 prf.2510326B Streptomyces peucetius dnrV 31.0 52.1 284 doxorubicin biosynthesis enzyme 168 3668 160029 160370 342 gp:SPAC1250_3 Schizosaccharomyces pombe 35.6 56.7 104 methyltransferase SPAC1250.04c 169 3669 160431 161360 930 170 3670 161696 162352 657 171 3671 162295 161363 933 172 3672 162463 162867 405 gp:AE002420_13 Neisseria meningitidis MC58 41.5 76.3 118 ribonuclease NMB0662 173 3673 162965 163603 639 174 3674 165717 166457 741 175 3675 165755 163689 2067 gp:AF176569_1 Mus musculus nl1 28.5 57.2 722 neprilysin-like metallopeptidase 1 176 3676 166457 167419 963 177 3677 168595 167837 759 sp:FARR_ECOLI Escherichia coli K12 farR 29.8 65.6 238 transcriptional regulator, GntR family or fatty acyl-responsive regulator 178 3678 168975 169991 1017 pir:T14544 Beta vulgaris 28.6 63.0 332 fructokinase or carbohydrate kinase 179 3679 169996 170916 921 gp:SC8F11_3 Streptomyces coelicolor A3(2) 52.7 80.7 296 hypothetical protein SC8F11.03c 180 3680 170933 172444 1512 prf:2204281A Streptomyces coelicolor msdA 61.0 86.1 498 methylmalonic acid semialdehyde dehydrogenase 181 3681 172468 173355 888 sp:IOLB_BACSU Bacillus subtilis iolB 33.2 58.2 268 myo-inositol catabolism 182 3682 173548 175275 1728 sp:IOLD_BACSU Bacillus subtilis iolD 41.0 69.8 586 myo-inositol catabolism 183 3683 175319 176272 954 sp:MOCC_RHIME Rhizobium meliloti mocC 29.7 51.0 290 rhizopine catabolism protein 184 3684 176308 177318 1011 sp:MI2D_BACSU Bacillus subtilis idh or iolG 39.1 72.2 335 myo-inositol 2-dehydrogenase 185 3685 177334 178203 870 sp:IOLH_BACSU Bacillus subtilis iolH 44.6 72.1 287 myo-inositol catabolism 186 3686 178285 179658 1374 sp:TCMA_STRGA Streptomyces glaucescens tcmA 30.9 61.5 457 metabolite export pump of tetracenomycin C resistance 187 3687 179081 178461 621 188 3688 179689 180711 1023 sp:YVAA_BACSU Bacillus subtilis yvaA 31.1 65.5 354 oxidoreductase 189 3689 180842 181297 456 190 3690 181264 181647 384 191 3691 182679 181687 993 gp:SRE9798_1 Streptomyces reticuli cebR 32.0 61.9 331 regulatory protein 192 3692 182819 184051 1233 sp:Y4HM_RHISN Rhizobium sp. NGR234 y4hM 24.4 52.5 442 oxidoreductase 193 3693 184077 185087 1011 sp:YFIH_BACSU Bacillus subtilis yfiH 33.7 64.7 303 hypothetical protein 194 3694 185214 185642 429 195 3695 186508 186708 201 sp:CSP_ARTGO Streptomyces coelicolor A3(2) 70.3 92.2 64 cold shock protein csp 196 3696 186769 187302 534 197 3697 187302 187607 306 198 3698 187687 188100 414 prf:2113413A Stellaria longipes 30.6 58.2 134 caffeoyl-CoA 3-O-methyltransferase 199 3699 188725 188300 426 200 3700 189736 188747 990 sp:CCPA_BACSU Bacillus subtilis ccpA 28.7 62.1 338 glucose-resistance amylase regulator regulator 201 3701 189920 190321 402 202 3702 190628 190389 240 203 3703 192175 190703 1473 sp:XYLT_LACBR Lactobacillus brevis xylT 36.0 70.5 458 D-xylose proton symporter 204 3704 193248 192949 300 205 3705 193262 194464 1203 gp:AF189147_1 Corynebacterium glutamicum 100.0 100.0 401 transposase (ISCg2) ATCC 13032 tnp 206 3706 195038 194604 435 sp:FIXL_RHIME Rhizobium meliloti fixL 27.6 60.7 145 signal-transducing histidine kinase 207 3707 195240 199769 4530 gp:AB024708_1 Corynebacterium glutamicum 99.9 100.0 1510 glutamine 2-oxoglutarate gltB aminotransferase large subunit 208 3708 199772 201289 1518 gp:AB024708_2 Corynebacterium glutamicum 99.4 99.8 506 glutamine 2-oxoglutarate gltD aminotransferase small subunit 209 3709 201580 201341 240 210 3710 203244 201760 1485 pir:C70793 Mycobacterium tuberculosis 44.6 72.8 496 hypothetical protein H37Rv Rv3698 211 3711 205588 205956 369 212 3712 206068 206385 318 213 3713 207011 203541 3471 prf:2224383C Mycobacterium avium embB 39.8 70.6 1122 arabinosyl transferase 214 3714 208989 207007 1983 pir:D70697 Mycobacterium tuberculosis 35.0 66.1 651 hypothetical membrane protein H37Rv Rv3792 215 3715 209968 209210 759 prf:2504279B Pseudomonas sp. phbB 31.4 56.5 223 acetoacetyl CoA reductase 216 3716 211455 209992 1464 pir:B70697 Mycobacterium tuberculosis 66.0 85.1 464 oxidoreductase H37Rv Rv3790 217 3717 211768 211535 234 218 3718 211777 212283 507 219 3719 212283 212735 453 220 3720 212656 213657 1002 gp:LMA243459_1 Leishmania major ppg1 24.3 57.4 350 proteophosphoglycan 221 3721 213712 214107 396 sp:YOGN_MYCTU Mycobacterium tuberculosis 60.5 83.9 124 hypothetical protein H37Rv Rv3789 222 3722 214121 214522 402 223 3723 214527 215159 633 pir:H70666 Mycobacterium tuberculosis 43.2 73.8 206 hypothetical protein H37Rv Rv1864c 224 3724 216100 215162 939 pir:B70696 Mycobacterium tuberculosis 63.6 79.1 302 rhamnosyl transferase H37Rv Rv3782 rfbE 225 3725 216264 216605 342 226 3726 216712 216116 597 gp:AB016260_100 Agrobacterium tumefaciens 313 55.1 214 hypothetical protein plasmid pTi-SAKURA tiorf100 227 3727 217929 217141 789 sp:RFBE_YEREN Yersinia enterocolitica rfbE 47.0 78.4 236 O-antigen export system ATP- binding protein 228 3728 218746 217943 804 sp:RFBD_YEREN Yersinia enterocolitica rfbD 31.3 75.6 262 O-antigen export system permease protein 229 3729 218979 220151 1173 pir:F70695 Mycobacterium tuberculosis 36.5 63.0 416 hypothetical protein H37Rv Rv3778c 230 3730 221107 220154 954 gp:AF010309_1 Homo sapiens pig3 41.1 71.5 302 NADPH quinone oxidoreductase 231 3731 221712 221131 582 232 3732 221911 222207 297 PIR:A70606 Mycobacterium tuberculosis 35.0 51.0 78 probable electron transfer protein H37Rv Rv3571 233 3733 223685 222210 1476 sp:ALST_BACSU Bacillus subtilis alsT 46.7 75.8 475 amino acid carrier protein 234 3734 224336 225244 909 235 3735 226324 225242 1083 gp:SYPCCMOEB_1 Synechococcus sp. PCC 7942 43.8 70.1 368 molybdopterin biosynthesis protein moeB moeB (sulfurylase) 236 3736 226767 226312 456 prf:2403296D Arthrobacter nicotinovorans 44.7 75.3 150 molybdopterin synthase, large moaE subunit 237 3737 227230 226760 471 sp:MOCB_SYNP7 Synechococcus sp. PCC 7942 33.5 63.3 158 molybdenum cofactor biosynthesis moaCB protein CB 238 3738 227685 227218 468 prf:2403296C Arthrobacter nicotinovorans 61.7 84.4 154 co-factor synthesis protein moaC 239 3739 228887 227703 1185 gp:ANY10817_2 Arthrobacter nicotinovorans 34.5 58.6 377 molybdopterin co-factor synthesis moeA protein 240 3740 229613 228891 723 prf:2403296F Arthrobacter nicotinovorans 44.1 70.5 227 hypothetical membrane protein modB 241 3741 230514 229711 804 prf:2403296E Arthrobacter nicotinovorans 34.0 68.0 256 molybdate-binding periplasmic modA protein 242 3742 230608 230928 321 pir:D70816 Mycobacterium tuberculosis 37.5 70.8 96 molybdopterin converting factor H37Rv moaD2 subunit 1 243 3743 231842 230931 912 prf:2518354A Thermococcus litoralis malK 34.3 60.8 365 maltose transport protein 244 3744 232267 231848 420 sp:YPT3_STRCO Streptomyces coelicolor A3(2) 36.4 76.9 121 hypothetical membrane protein ORF3 245 3745 233282 232260 1023 sp:HIS8_ZYMMO Zymomonas mobilis hisC 37.3 65.8 330 histidinol-phosphate aminotransferase 246 3746 233913 234818 906 247 3747 235203 234910 294 248 3748 235290 235409 120 249 3749 236212 235451 762 gp:BAU81286_1 Brucella abortus oxyR 29.4 57.1 252 transcription factor 250 3750 236326 237342 1017 sp:ADH2_BACST Bacillus stearothermophilus 34.0 66.0 335 alcohol dehydrogenase DSM 2334 adh 251 3751 237345 238145 801 sp:PUO_MICRU Micrococcus rubens puo 21.5 38.1 451 putrescine oxidase 252 3752 238176 239525 1350 prf:2305239A Borrelia burgdorferi mgtE 30.9 68.5 444 magnesium ion transporter 253 3753 239772 239945 174 254 3754 239986 241515 1530 prf:2320140A Xenopus laevis 33.2 59.6 567 Na/dicarboxylate cotransporter 255 3755 242902 241883 1020 pir:C70800 Mycobacterium tuberculosis 46.1 69.1 317 oxidoreductase H37Rv tyrA 256 3756 242910 243431 522 pir:B70800 Mycobacterium tuberculosis 48.8 73.8 160 hypothetical protein H37Rv Rv3753c 257 3757 243494 243910 417 gp:RHBNFXP_1 Bradyrhizobium japonicum 45.1 70.1 144 nitrogen fixation protein 258 3758 244015 244215 201 259 3759 244466 244816 351 260 3760 244902 247304 2403 sp:YV34_MYCTU Mycobacterium tuberculosis 20.7 45.7 997 membrane transport protein H37Rv Rv0507 mmpL2 261 3761 247310 248572 1263 sp:TGT_ZYMMO Zymomonas mobilis 41.3 68.0 400 queuine tRNA-ribosyltransferase 262 3762 249294 248557 738 sp:YPDP_BACSU Bacillus subtilis ypdP 28.1 62.1 203 hypothetical membrane protein 263 3763 249428 250507 1080 264 3764 250369 249722 648 265 3765 250503 251939 1437 pir:S65588 Streptomyces glaucescens strW 24.3 49.6 526 ABC transporter 266 3766 251952 252830 879 sp:SYE_BACSU Bacillus subtilis gltX 34.8 63.3 316 glutamyl-tRNA synthetase 267 3767 253819 252830 990 268 3768 255438 254329 1110 gp:PSESTBCBAD_1 Pseudomonas syringae tnpA 34.2 55.0 360 transposase 269 3769 255794 255492 303 270 3770 256067 256204 138 271 3771 256599 257894 1296 gsp:W69554 Brevibacterium lactofermentum 98.6 100.0 432 aspartate transaminase aspC 272 3772 257900 258529 630 273 3773 258551 260875 2325 gp:AF025391_1 Thermus thermophilus dnaX 31.6 53.1 642 DNA polymerase III holoenzyme tau subunit 274 3774 259312 258596 717 275 3775 260987 261295 309 sp:YAAK_BACSU Bacillus subtilis yaaK 41.6 74.3 101 hypothetical protein 276 3776 261402 262055 654 sp:RECR_BACSU Bacillus subtilis recR 42.5 72.4 214 recombination protein 277 3777 263295 262546 750 prf:2503462B Heliobacillus mobilis cobQ 38.3 61.7 248 cobyric acid synthase 278 3778 264566 263298 1269 prf:2503462C Heliobacillus mobilis murC 31.3 60.6 444 UDP-N-acetylmuramyl tripeptide synthetase 279 3779 265678 264599 1080 pir:H70794 Mycobacterium tuberculosis 25.7 55.2 346 DNA polymerase III epsilon chain H37Rv dnaQ 280 3780 269124 268258 867 sp:YLEU_CORGL Corynebacterium glutamicum 100.0 100.0 270 hypothetical membrane protein (Brevibacterium flavum) ATCC 13032 orfX 281 3781 269371 270633 1263 sp:AKAB_CORGL Corynebacterium glutamicum 99.5 99.8 421 aspartate kinase alpha chain lysC-alpha 282 3782 270576 269524 1053 283 3783 271761 273194 1434 284 3784 274120 273542 579 prf:2312309A Mycobacterium smegmatis sigE 31.2 63.5 189 extracytoplasmic function alternative sigma factor 285 3785 274366 275871 1506 sp:CATV_BACSU Bacillus subtilis katA 52.9 76.4 492 vegetative catalase 286 3786 275891 276232 342 287 3787 276247 275957 291 288 3788 276763 276302 462 sp:LRP_KLEPN Klebsiella pneumoniae lrp 37.1 72.0 143 leucine-responsive regulatory protein 289 3789 276829 277581 753 sp:AZLC_BACSU Bacillus subtilis 1A1 azlC 30.5 68.0 203 branched-chain amino acid transport 290 3790 277581 277904 324 291 3791 278301 277987 315 292 3792 278732 278388 345 gp:AF178758_1 Sinorhizobium sp. As4 arsR 34.4 68.9 90 metalloregulatory protein 293 3793 278814 279893 1080 gp:AF178758_2 Sinorhizobium sp. As4 arsB 52.2 84.2 341 arsenic oxyanion-translocation pump membrane subunit 294 3794 279893 280279 387 sp:ARSC_STAXY Staphylococcus xylosus arsC 31.1 68.9 119 arsenate reductase 295 3795 280666 280349 318 296 3796 280939 280670 270 297 3797 281401 280949 453 298 3798 282933 281404 1530 gp:AF097740_4 Bacillus firmus OF4 mrpD 32.4 70.4 503 Na+/H+ antiporter or multiple resistance and pH regulation related protein D 299 3799 283317 282937 381 prf:2504285D Staphylococcus aureus mnhC 37.0 70.6 119 Na+/H+ antiporter 300 3800 286202 283317 2886 gp:AF097740_1 Bacillus firmus OF4 mrpA 34.1 64.3 824 Na+/H+ antiporter or multiple resistance and pH regulation related protein A 301 3801 286373 287857 1485 302 3802 287661 287059 603 303 3803 288829 287966 864 304 3804 289796 289131 666 sp:CZCR_ALCEU Alcaligenes eutrophus CH34 38.6 70.4 223 transcriptional activator czcR 305 3805 291243 289777 1467 prf:2214304B Mycobacterium tuberculosis 26.7 56.8 521 two-component system sensor mtrB histidine kinase 306 3806 291815 292417 603 sp:APL_LACLA Lactococcus lactis MG1363 apl 28.3 60.0 180 alkaline phosphatase 307 3807 291833 291273 561 308 3808 293511 292597 915 pir:B69865 Bacillus subtilis ykuE 26.1 54.7 307 phosphoesterase 309 3809 293539 293991 453 sp:YQEY_BACSU Bacillus subtilis yqeY 37.6 71.8 149 hypothetical protein 310 3810 296388 294004 2385 prf:2209359A Mycobacterium leprae pon1 48.3 77.1 782 class A penicillin-binding protein(PBP1) 311 3811 297064 297402 339 pir:S20912 Streptomyces coelicolor A3(2) 40.9 63.4 71 regulatory protein whiB 312 3812 297431 297622 192 313 3813 297631 297783 153 gp:SCH17_10 Streptomyces coelicolor A3(2) 84.0 96.0 50 hypothetical protein SCH17.10c 314 3814 297792 298250 459 pir:G70790 Mycobacterium tuberculosis 65.1 89.9 149 transcriptional regulator H37Rv Rv3678c 315 3815 299684 298332 1353 sp:SHIA_ECOLI Escherichia coli K12 shiA 37.3 68.9 440 shikimate transport protein 316 3816 300087 300695 609 317 3817 301261 299726 1536 sp:LCFA_BACSU Bacillus subtilis lcfA 31.1 59.9 534 long-chain-fatty-acid-CoA ligase 318 3818 302036 301512 525 gp:SCJ4_28 Streptomyces coelicolor A3(2) 33.9 65.4 127 transcriptional regulator SCJ4.28c 319 3819 302167 303099 933 sp:FABG_BACSU Bacillus subtilis fabG 41.0 72.5 251 3-oxoacyl-(acyl-carrier-protein) reductase 320 3820 303133 304074 942 sp:FLUG_EMENI Emericella nidulans fluG 27.2 52.0 254 glutamine synthetase 321 3821 304070 305263 1194 prf:2512386A Arabidopsis thaliana atg6 38.8 66.5 394 short-chain acyl CoA oxidase 322 3822 305288 305758 471 sp:NODN_RHILV Rhizobium leguminosarum nodN 45.8 72.6 153 nodulation protein 323 3823 305858 306700 843 pir:F70790 Mycobacterium tuberculosis 41.2 72.4 272 hydrolase H37Rv Rv3677c 324 3824 306367 305195 1173 325 3825 306800 307504 705 326 3826 307462 306782 681 prf.2323349A Vibrio cholerae crp 30.9 65.7 207 cAMP receptor protein 327 3827 307918 307727 192 328 3828 307955 308734 780 sp:UVEN_MICLU Micrococcus luteus pdg 57.5 77.1 240 ultraviolet N-glycosylase/AP lyase 329 3829 308745 309302 558 pirB70790 Mycobacterium tuberculosis 34.6 58.3 211 cytochrome c biogenesis protein H37Rv Rv3673c 330 3830 309370 310038 669 sp:YEAB_ECOLI Escherichia coli K12 yeaB 30.7 56.3 192 hypothetical protein 331 3831 310135 311325 1191 pir:H70789 Mycobacterium tuberculosis 38.6 71.0 396 serine proteinase H37Rv Rv3671c 332 3832 312891 311899 993 prf:2411250A Corynebacterium sp. C12 cEH 29.6 52.1 280 epoxide hydrolase 333 3833 313457 312909 549 pir: F70789 Mycobacterium tuberculosis 46.8 77.6 156 hypothetical membrane protein 334 3834 314590 313625 966 pir:S72914 Mycobacterium leprae 29.6 65.5 287 phosphoserine phosphatase MTCY20G9.32C. serB 335 3835 314980 316002 1023 pir:E70788 Mycobacterium tuberculosis 35.0 60.2 349 hypothetical protein H37Rv Rv3660c 336 3836 316110 317132 1023 pir:C44020 Escherichia coli trbB 32.9 66.5 319 conjugal transfer region protein 337 3837 316964 316350 615 338 3838 317078 317893 816 pir:C70788 Mycobacterium tuberculosis 30.5 63.7 262 hypothetical membrane protein H37Rv Rv3658c 339 3839 317920 318465 546 pir:B70788 Mycobacterium tuberculosis 33.8 64.2 201 hypothetical protein H37Rv Rv3657c 340 3840 318492 318689 198 pir:A70788 Mycobacterium tuberculosis 47.5 84.8 59 hypothetical protein H37Rv Rv3656c 341 3841 318696 319013 318 342 3842 318958 318545 414 343 3843 318991 319335 345 344 3844 321690 319336 2355 sp:YPRA_BACSU Bacillus subtilis yprA 33.8 66.1 764 ATP-dependent RNA helicase 345 3845 322007 322207 201 sp:CSP_ARTGO Arthrobacter globiformis Sl55 68.7 88.1 67 cold shock protein csp 346 3846 322216 321992 225 347 3847 322910 325897 2988 pir:G70563 Mycobacterium tuberculosis 61.7 81.6 977 DNA topoisomerase I H37Rv Rv3646c topA 348 3848 325904 326614 711 349 3849 327735 326695 1041 sp:CYAB_STIAU Stigmatella aurantiaca B17R20 32.7 62.4 263 adenylate cyclase cyaB 350 3850 328283 329539 1257 sp:DP3X_BACSU Bacillus subtilis dnaX 25.3 52.7 423 DNA polymerase III subunit tau/gamma 351 3851 329748 329909 162 352 3852 329933 330376 444 gp:AE002103_3 Ureaplasma urealyticum uu033 32.6 59.0 144 hypothetical protein 353 3853 330973 331533 561 gp:AE001882_8 Deinococcus radiodurans 39.0 63.4 172 hypothetical protein DR0202 354 3854 331552 332433 882 sp:RLUC_ECOLI Escherichia coli K12 rluC 43.6 65.0 314 ribosomal large subunit pseudouridine synthase C 355 3855 332919 334562 1644 sp:BGLX_ERWCH Erwinia chrysanthemi D1 bgxA 34.8 60.2 558 beta-glucosidase/xylosidase 356 3856 332965 334953 1989 gp:AF090429_2 Azospirillum irakense salB 38.6 61.4 101 beta-glucosidase 357 3857 335009 336112 1104 sp:FADH_AMYME Amycolatopsis methanolica 66.6 86.5 362 NAD/mycothiol-dependent formaldehyde dehydrogenase 358 3858 335805 335185 621 359 3859 336212 336748 537 sp:YTH5_RHOSN Rhodococcus erythropolis orf5 32.5 47.5 160 metallo-beta-lactamase superfamily 360 3860 336781 337449 669 sp:FABG_ECOLI Escherichia coli K12 fabG 25.9 55.8 251 3-oxoacyl-(acyl-carrier-protein) reductase 361 3861 337539 338768 1230 gp:AF148322_1 Streptomyces viridifaciens vlmF 26.3 56.4 415 valanimycin resistant protein 362 3862 338793 339725 933 prf:2512357B Actinoplanes sp. acbB 33.8 66.3 320 dTDP-glucose 4,6-dehydratase 363 3863 340569 340195 375 pir:A70562 Mycobacterium tuberculosis 59.3 88.9 108 hypothetical protein H37Rv Rv3632 364 3864 341327 340569 759 sp:YC22_METJA Methanococcus jannaschii JAL- 33.9 66.5 230 dolichol phosphate mannose 1 MJ1222 synthase 365 3865 341347 342375 1029 366 3866 342417 343451 1035 sp:YEFJ_ECOLI Escherichia coli K12 yefJ 25.8 57.3 260 nucleotide sugar synthetase 367 3867 343636 345717 2082 sp:USHA_SALTY Salmonella typhimurium ushA 26.1 54.4 586 UDP-sugar hydrolase 368 3868 345975 345814 162 369 3869 346460 346110 351 370 3870 348019 346961 1059 sp:ADH_MYCTU Mycobacterium tuberculosis 52.2 74.9 343 NADP-dependent alcohol H37Rv adhC dehydrogenase 371 3871 348952 348098 855 sp:RFBA_SALAN Salmonella anatum M32 rfbA 62.8 84.9 285 glucose-1-phosphate thymidylyltransferase 372 3872 350310 348952 1359 gp:D78182_5 Streptococcus mutans rmlC 49.5 74.0 192 dTDP-4-keto-L-rhamnose reductase 373 3873 351443 350313 1131 sp:RMLB_STRMU Streptococcus mutans XC rmlB 61.8 83.4 343 dTDP-glucose 4,6-dehydratase 374 3874 351948 351370 579 sp:NOX_THETH Thermus aquaticus HB8 nox 35.4 61.2 206 NADH dehydrogenase 375 3875 352693 353637 945 prf:2510361A Staphylococcus aureus sirA 33.2 66.5 325 Fe-regulated protein 376 3876 354387 353749 639 377 3877 355906 354599 1308 sp:Y17M_MYCTU Mycobacterium tuberculosis 37.4 68.3 423 hypothetical membrane protein H37Rv Rv3630 378 3878 357228 355849 1380 gp:SC5F2A_19 Streptomyces coelicolor 34.1 62.5 461 metallopeptidase SC5F2A.19c 379 3879 359354 357237 2118 prf:2502226A Sphingomonas capsulata 28.4 56.4 708 prolyl endopeptidase 380 3880 360334 359762 573 381 3881 361905 360814 1092 gp:SCF43_2 Streptomyces coelicolor A3(2) 26.0 46.0 258 hypothetical membrane protein 382 3882 363151 362057 1095 gsp:W56155 Corynebacterium 50.7 76.6 363 cell surface layer protein ammoniagenes ATCC 6872 383 3883 363824 365257 1434 prf:2404346B Acinetobacter johnsonii ptk 28.5 57.2 453 autophosphorylating protein Tyr kinase 384 3884 365250 365852 603 prf:2404346A Acinetobacter johnsonii ptp 39.2 68.6 102 protein phosphatase 385 3885 365855 366838 984 386 3886 366832 368643 1812 sp:CAPD_STAAU Staphylococcus aureus M capD 33.0 65.7 613 capsular polysaccharide biosynthesis 387 3887 368642 367701 942 PRF:2109288X Vibrio cholerae 41.0 51.0 90 ORF 3 388 3888 368647 369801 1155 prf:2423410L Campylobacter jejuni wlaK 37.1 68.3 394 lipopolysaccharide biosynthesis/ aminotransferase 389 3889 369794 370405 612 gp:AF014804_1 Neisseria meningitidis pglB 54.6 75.0 196 pilin glycosylation protein 390 3890 370613 371773 1161 sp:CAPM_STAAU Staphylococcus aureus M capM 33.4 69.2 380 capsular polysaccharide biosynthesis 391 3891 371929 373419 1491 pir:S67859 Xanthornonas campestris gumJ 34.3 69.8 504 lipopolysacoharide biosynthesis/ export protein 392 3892 373500 374813 1314 sp:MURA_ENTCL Enterobacter cloacae murA 31.4 64.6 427 UDP-N-acetylglucosamine 1- carboxyvinyltransferase 393 3893 374833 375837 1005 sp:MURB_BACSU Bacillus subtilis murB 34.8 68.5 273 UDP-N-acetylenolpyruvoylglucosamine reductase 394 3894 375842 376878 1035 gp:VCLPSS_9 Vibrio cholerae ORF39x2 32.0 57.3 356 sugar transferase 395 3895 377683 377832 150 prf:2211295A Corynebacterium glutamicum 60.4 79.3 53 transposase 396 3896 378093 378227 135 397 3897 378185 378511 327 pir:S43613 Corynebacterium glutamicum 75.7 94.3 70 transposase (insertion sequence ATCC 31831 IS31831) 398 3898 378562 378287 276 399 3899 379837 378668 1170 pir:G70539 Mycobacteriurn tuberculosis 28.0 57.4 404 hypothetical protein H37Rv Rv1565c 400 3900 380842 379850 993 gsp:W37352 Pseudomonas aeruginosa PAO1 34.5 60.2 354 acetyltransferase psbC 401 3901 381265 381495 231 PIR:S60890 Corynebacterium glutamicum 44.0 53.0 65 hypothetical protein B 402 3902 381948 383108 1161 sp:UDG8_ECOLI Escherichia coli ugd 63.7 89.7 388 UDP-glucose 6-dehydrogenase 403 3903 383768 383496 273 404 3904 385190 383982 1209 405 3905 386195 385374 822 gp:AF172324_3 Escherichia coli wbnA 32.1 65.0 243 glycosyl transferase 406 3906 386556 387200 645 gp:A2008676_13 Escherichia coli 0157 wbhH 33.0 62.0 221 acetyltransferase 407 3907 387657 387463 195 408 3908 387692 389098 1407 gp:CGLPD_1 Corynebacterium glutamicum 99.6 100.0 469 dihydrolipoamide dehydrogenase ATCC 13032 Ipd 409 3909 389248 390168 921 pir:JC4985 Xanthomonas campestris 41.7 68.1 295 UTP-glucose-1-phosphate uridylyltransferase 410 3910 390233 390730 498 gp:PAU49666_2 Pseudomonas aeruginosa PAO1 43.8 71.9 153 regulatory protein orfX 411 3911 392208 390787 1422 pir:E70828 Mycobacterium tuberculosis 57.0 81.3 477 transcriptional regulator H37Rv Rv0465c 412 3912 392705 393475 771 gp:SCM10_12 Streptomyces coelicolor A3(2) 34.8 67.4 230 cytochrome b subunit SCM10.12c 413 3913 393639 395513 1875 pir:A27763 Bacillus subtilis sdhA 32.4 61.2 608 succinate dehydrogenase flavoprotein 414 3914 395426 396262 837 gp:BMSDHCAB_4 Paenibacillus macerans sdhB 27.5 56.2 258 succinate dehydrogenase subunit B 415 3915 396315 396650 336 416 3916 396672 396932 261 417 3917 397040 396411 630 418 3918 397730 397825 96 419 3919 397884 398222 339 420 3920 398206 397232 975 gp:SCC78_5 Streptomyces coelicolor 26.3 49.8 259 hypothetical protein SCC78.05 421 3921 398329 399579 1251 sp:YJIN_ECOLI Escherichia coli K12 yjiN 32.7 64.3 431 hypothetical protein 422 3922 399598 400017 420 423 3923 400039 400341 303 424 3924 400473 401150 678 sp:TCMR_STRGA Streptomyces glaucescens 26.4 53.8 197 tetracenomycin C transcription GLA.0 tcmR repressor 425 3925 401050 401253 204 426 3926 401150 402796 1647 gp:AF164961_8 Streptomyces fradiae T#2717 36.1 74.6 499 transporter urdJ 427 3927 402799 404430 1632 gp:AF164961_8 Streptomyces fradiae T#2717 39.6 74.6 508 transporter urdJ 428 3928 405419 404508 912 sp:PURU_CORSP Corynebacterium sp. P-1 purU 40.9 72.7 286 formyltetrahydrofolate deformylase 429 3929 405480 406145 666 sp.DEOC_BACSU Bacillus subtilis deoC 38.5 74.0 208 deoxyribose-phosphate aldolase 430 3930 406310 406161 150 431 3931 406417 405521 897 432 3932 406550 407416 867 prf:2413441K Mycobacterium avium GIR10 26.8 53.6 280 hypothetical protein mav346 433 3933 407708 407409 300 pir:A70907 Mycobacterium tuberculosis 58.7 85.9 92 hypothetical protein H37Rv Rv0190 434 3934 408546 409145 600 435 3935 409975 407711 2265 sp:CTPB_MYCLE Mycobacterium leprae ctpB 45.7 75.3 748 cation-transporting P-type ATPase B 436 3936 410476 410027 450 437 3937 410683 412545 1863 sp:AMYH_YEAST Saccharomyces cerevisiae 27.3 56.1 626 glucan 1,4-alpha-glucosidase S288C YIR019C sta1 438 3938 412557 413633 1077 gp:AF109162_1 Corynebacterium diphtheriae 57.2 83.6 348 hemin-binding periplasmic protein hmuT 439 3939 413643 414710 1068 gp:AF109162_2 Corynebacterium diphtheriae 65.2 90.3 330 ABC transporter hmuU 440 3940 414714 415526 813 gp:AF109162_3 Corynebacterium diphtheriae 63.8 85.0 254 ABC transporter ATP-binding protein hmuV 441 3941 415643 416599 957 gp:SCC75A_17 Streptomyces coelicolor C75A 28.6 56.4 266 hypothetical protein SCC75A.17c 442 3942 416603 417439 837 gp:SCC75A_17 Streptomyces coelicolor C75A 32.6 61.6 258 hypothetical protein SCC75A.17c 443 3943 418354 417545 810 444 3944 419253 418441 813 445 3945 419757 419257 501 446 3946 419785 420885 1101 gp:ECOMURBA_1 Escherichia coli RDD012 murB 30.1 58.4 356 UDP-N-acetylpyruvoylglucosamine reductase 447 3947 420866 421516 651 448 3948 421043 420309 735 449 3949 421858 422031 174 450 3950 423793 422090 1704 sp:LCFA_BACSU Bacillus subtilis lcfA 35.5 68.1 558 long-chain-fatty-acid-CoA ligase 451 3951 423878 425131 1254 gp:SC2G5_6 Streptomyces coelicolor 33.9 58.7 416 transferase SC2G5.06 452 3952 425177 425920 744 sp:PMGY_STRCO Streptomyces coelicolor A3(2) 70.7 84.2 246 phosphoglycerate mutase gpm 453 3953 425934 427172 1239 prf:2404434A Mycobacterium bovis senX3 49.2 74.8 417 two-component system sensor histidine kinase 454 3954 427172 427867 696 prf:2404434B Mycobacterium bovis BCG 75.8 90.9 231 two-component response regulator regX3 455 3955 428561 429439 879 456 3956 432023 429438 2586 gp:SCE25_30 Streptomyces coelicolor A3(2) 31.3 60.7 921 ABC transporter ATP-binding protein SCE25.30 457 3957 433028 432126 903 sp:YV21_MYCTU Mycobacterium tuberculosis 45.0 66.9 269 cytochrome P450 H37Rv RV3121 458 3958 433062 433988 927 prf:2512277A Pseudomonas aeruginosa ppx 28.8 57.8 306 exopolyphosphatase 459 3959 434010 434822 813 sp:YV23_MYCTU Mycobacterium tuberculosis 28.8 57.3 302 hypothetical membrane protein H37Rv Rv0497 460 3960 434886 435695 810 sp:PROC_CORGL Corynebacterium glutamicum 100.0 100.0 269 pyrroline-5-carboxylate reductase ATCC 17965 proC 461 3961 434986 433865 1122 gp:D88733_1 Equine herpesvirus 1 ORF71 25.4 52.0 394 membrane glycoprotein 462 3962 435940 436137 198 pir:S72921 Mycobacterium leprae 76.4 94.6 55 hypothetical protein B2168_C1_172 463 3963 436321 436103 219 464 3964 436463 436561 99 gp:SCE68_25 Streptomyces coelicolor 89.7 100.0 29 hypothetical protein SCE68.25c 465 3965 436573 436764 192 466 3966 437233 437850 618 467 3967 438044 436980 1065 pir:S72914 Mycobacterium leprae 51.0 77.4 296 phosphoserine phosphatase MTCY20G9.32C.serB 468 3968 438179 438424 246 sp:YV35_MYCTU Mycobacterium tuberculosis 40.5 66.2 74 hypothetical protein H37Rv Rv0508 469 3969 438294 438037 258 470 3970 438516 439904 1389 sp:HEM1_MYCLE Mycobacterium leprae hemA 44.4 74.3 455 glutamyl-tRNA reductase 471 3971 439909 440814 906 pir:S72887 Mycobacterium leprae hem3b 50.7 75.3 308 hydroxymethylbilane synthase 472 3972 441220 441591 372 473 3973 442482 441601 882 sp:CATM_ACICA Acinetobacter calcoaceticus 27.1 57.6 321 cat operon transcriptional regulator catM 474 3974 442758 444158 1401 sp:SHIA_ECOLI Escherichia coli K12 shiA 35.5 72.2 417 shikimate transport protein 475 3975 444185 446038 1854 sp:3SHD_NEUCR Neurospora crassa qa4 28.2 57.9 309 3-dehydroshikimate dehydratase 476 3976 446538 447386 849 gp:AF124518_2 Corynebacterium glutamicum 98.2 98.6 282 shikimate dehydrogenase ASO19 aroE 477 3977 447670 447398 273 478 3978 449179 448130 1050 sp:POTG_ECOLI Escherichia coli K12 potG 34.7 68.6 363 putrescine transport protein 479 3979 449714 449100 615 480 3980 450826 449183 1644 sp:SFUB_SERMA Serratia marcescens sfuB 25.1 55.2 578 iron(III)-transport system permease protein 481 3981 450849 451961 1113 482 3982 451895 450837 1059 gp:SHU75349_1 Brachyspira hyodysenteriae bitA 25.1 59.9 347 periplasmic-iron-binding protein 483 3983 452661 454430 1770 pir:S72909 Mycobacterium leprae cysG 46.5 71.6 486 uroporphyrin-III C-methyltransferase 484 3984 454450 454875 426 485 3985 454967 455983 1017 sp:HEM2_STRCO Streptomyces coelicolor A3(2) 60.8 83.1 337 delta-aminolevulinic acid hemB dehydratase 486 3986 456016 456597 582 487 3987 456641 457150 510 488 3988 457357 459900 2544 sp:CTPB_MYCLE Mycobacterium leprae ctpB 27.4 56.5 858 cation-transporting P-type ATPase B 489 3989 459425 458583 843 490 3990 460020 461093 1074 sp:DCUP_STRCO Streptomyces coelicolor A3(2) 55.0 76.7 364 uroporphyrinogen decarboxylase hemE 491 3991 461112 462455 1344 sp:PPOX_BACSU Bacillus subtilis hemY 28.0 59.9 464 protoporphyrinogen IX oxidase 492 3992 462557 463867 1311 sp:GSA_MYCLE Mycobacterium leprae hemL 61.7 83.5 425 glutamate-1-semialdehyde 2,1- aminomutase 493 3993 463867 464472 606 sp:PMG2_ECOLI Escherichia coli K12 gpmB 28.0 62.7 161 phosphoglycerate mutase 494 3994 464482 465102 621 pir:A70545 Mycobacterium tuberculosis 44.7 71.2 208 hypothetical protein H37Rv Rv0526 495 3995 465118 465909 792 pir:B70545 Mycobacterium tuberculosis 53.5 85.3 245 cytochrome c-type biogenesis H37Rv ccsA protein 496 3996 465949 467571 1623 pir:C70545 Mycobacterium tuberculosis 50.7 76.0 533 hypothetical membrane protein H37Rv Rv0528 497 3997 467648 468658 1011 pir:070545 Mycobacterium tuberculosis 44.1 77.8 338 cytochrome c biogenesis protein H37Rv ccsB 498 3998 469370 470170 801 499 3999 470184 470654 471 pir:G70790 Mycobacterium tuberculosis 38.9 69.4 144 transcriptional regulator H37Rv Rv3678c pb5 500 4000 471013 470657 357 prf:2420312A Staphylococcus aureus zntR 31.1 72.2 90 Zn/Co transport repressor 501 4001 471420 471121 300 502 4002 471515 471847 333 pir:F70545 Mycobacterium tuberculosis 39.0 78.1 82 hypothetical membrane protein H37Rv Rv0531 503 4003 472808 471915 894 sp:MENA_ECOLI Escherichia coli K12 menA 33.6 61.5 301 1,4-dihydroxy-2-naphthoate octaprenyltransferase 504 4004 472948 473811 864 gp:AF125164_6 Bacteroides fragilis wcgB 32.4 62.6 238 glycosyl transferase 505 4005 475136 473814 1323 prf:2423270B Rhizobium trifolii matB 25.4 51.5 421 malonyl-CoA-decarboxylase 506 4006 475407 474997 411 sp:YQJF_ECOLI Escherichia coli K12 yqjF 35.3 65.5 139 hypothetical membrane protein 507 4007 477048 475489 1560 pir:S27612 Pseudomonas putida 50.4 76.0 520 ketoglutarate semialdehyde dehydrogenase 508 4008 477995 477048 948 sp:KDGD_PSEPU Pseudomonas putida KDGDH 48.5 75.6 303 5-dehydro-4-deoxyglucarate dehydratase 509 4009 478970 478092 879 sp:ALSR_BACSU Bacillus subtilis 168 alsR 36.9 66.2 293 als operon regulatory protein 510 4010 479303 478989 315 pir:B70547 Mycobacterium tuberculosis 33.0 64.9 94 hypothetical protein H37Rv Rv0543c 511 4011 480154 480597 444 512 4012 480201 479452 750 gp:SSP277295_9 Sphingomonas sp. LB126 fldB 28.1 54.7 267 2-pyrone-4,6-dicarboxylic acid 513 4013 480624 480208 417 514 4014 481001 480624 378 515 4015 481391 481131 261 516 4016 482668 481394 1275 pir:D70547 Mycobacterium tuberculosis 60.0 83.2 410 low-affinity inorganic phosphate H37Rv pitA transporter 517 4017 483587 483366 222 518 4018 483942 483637 306 519 4019 485062 484106 957 sp:MENB_BACSU Bacillus subtilis menB 48.5 70.3 293 naphthoate synthase 520 4020 485384 485986 603 gp:AE001957_12 Deinococcus radiodurans 57.9 82.7 202 peptidase E DR1070 521 4021 485385 485077 309 pir:C70304 Aquifex aeolicus VF5 phhB 37.7 68.8 77 pterin-4a-carbinolamine dehydratase 522 4022 486001 487014 1014 pir:D70548 Mycobacterium tuberculosis 54.0 76.7 335 muconate cycloisomerase H37Rv Rv0553 menC 523 4023 487028 488656 1629 sp:MEND_BACSU Bacillus subtilis menD 29.4 54.0 606 2-oxoglutarate decarboxylase and 2- succinyl-6-hydroxy-2,4- cyclohexadiene-1-carboxylate synthase 524 4024 488660 489100 441 pir:G70548 Mycobacterium tuberculosis 37.2 64.9 148 hypothetical membrane protein H37Rv Rv0556 525 4025 489209 490447 1239 pir:H70548 Mycobacterium tuberculosis 22.8 54.2 408 alpha-D-mannose-alpha(1- H37Rv pimB 6)phosphatidyl myo-inositol monomannoside transferase 526 4026 490580 491938 1359 sp:CYCA_ECOLI Escherichia coli K12 cycA 66.2 89.9 447 D-serine/D-alanine/glycine transporter 527 4027 491966 492655 690 sp:UBIE_ECOLI Escherichia coli K12 ubiE 37.1 66.7 237 ubiquinone/menaquinone biosynthesis methyltransferase 528 4028 492915 493583 669 529 4029 493916 492645 1272 pir:D70549 Mycobacterium tuberculosis 49.0 76.7 412 oxidoreductase H37Rv Rv0561c 530 4030 494061 495110 1050 sp:HEP2_BACST Bacillus stearothermophilus 39.2 67.1 316 heptaprenyl diphosphate synthase ATCC 10149 hepT component II 531 4031 496810 497142 333 gp:AF130462_2 Corynebacterium glutamicum 100.0 100.0 111 preprotein translocase SecE subunit ATCC 13032 secE 532 4032 497374 498327 954 gp:AF130462_3 Corynebacterium glutamicum 100.0 100.0 318 transcriptional antiterminator protein ATCC 13032 nusG 533 4033 498598 499032 435 gp:AF130462_4 Corynebacterium glutamicum 100.0 100.0 145 50S ribosomal protein L11 ATCC 13032 rplK 534 4034 499162 499869 708 gp:AF130462_5 Corynebacterium glutamicum 100.0 100.0 236 50S ribosomal protein L1 ATCC 13032 rplA 535 4035 501436 499925 1512 gp:SC5H4_2 Streptomyces coelicolor 23.1 50.2 564 regulatory protein SC5H4.02 536 4036 501577 502920 1344 sp:GABT_MYCTU Mycobacterium tuberculosis 60.5 82.4 443 4-aminobutyrate aminotransferase H37Rv RV2589 gabT 537 4037 502925 504283 1359 sp:GABD_ECOLI Escherichia coli K12 gabD 40.8 71.8 461 succinate-semialdehyde dehydrogenase (NAD(P)+) 538 4038 503739 503272 468 GP:ABCARRA_2 Azospirillum brasilense carR 32.0 38.0 150 novel two-component regulatory system 539 4039 504379 505569 1191 sp:TYRP_ECOLI Escherichia coli K12 o341#7 25.5 49.9 447 tyrosine-specific transport protein tyrP 540 4040 505698 507647 1950 sp:CTPG_MYCTU Mycobacterium tuberculosis 33.2 64.4 615 cation-transporting ATPase G H37Rv RV1992C ctpG 541 4041 507669 509081 1413 sp:P49_STRLI Streptomyces lividans P49 40.2 66.2 468 hypothetical protein or dehydrogenase 542 4042 509094 509696 603 543 4043 509998 510510 513 sp:RL10_STRGR Streptomyces griseus N2-3-11 52.9 84.7 170 50S ribosomal protein L10 rplJ 544 4044 510591 510974 384 sp:RL7_MYCTU Mycobacterium tuberculosis 72.3 89.2 130 50S ribosomal protein L7/L12 H37Rv RV0652 rplL 545 4045 511126 510989 138 546 4046 511536 512507 972 pir:A70962 Mycobacterium tuberculosis 25.8 55.5 283 hypothetical membrane protein H37Rv Rv0227c 547 4047 512913 516407 3495 sp:RPOB_MYCTU Mycobacterium tuberculosis 75.4 90.4 1180 DNA-directed RNA polymerase beta H37Rv RV0667 rpoB chain 548 4048 516494 520492 3999 sp:RPOC_MYCTU Mycobacterium tuberculosis 72.9 88.7 1332 DNA-directed RNA polymerase beta H37Rv RV0668 rpoC chain 549 4049 519277 518696 582 GP:AF121004_1 Mycobacterium tuberculosis 39.0 52.0 169 hypothetical protein H37Rv Jv0166c 550 4050 520671 520850 180 551 4051 520865 521644 780 gp:SCJ9A_15 Streptomyces coelicolor A3(2) 39.2 63.8 232 DNA-binding protein SCJ9A.15c 552 4052 522476 521679 798 sp:YT08_MYCTU Mycobacterium tuberculosis 29.3 57.7 215 hypothetical protein H37Rv RV2908C 553 4053 522694 523059 366 sp:RS12_MYCIT Mycobacterium intracellulare 90.9 97.5 121 30S ribosomal protein S12 rpsL 554 4054 523069 523533 465 sp:RS7_MYCSM Mycobacterium smegmatis 81.8 94.8 154 30S ribosomal protein S7 LR222 rpsG 555 4055 523896 526010 2115 sp:EFG_MICLU Micrococcus luteus fusA 71.7 88.9 709 elongation factor G 556 4056 526070 523911 2160 557 4057 526156 526013 144 558 4058 527121 526894 228 GSP:Y37841 Chlamydia trachomatis 56.0 78.0 44 lipoprotein 559 4059 527759 527607 153 560 4060 528040 528768 729 561 4061 529570 528779 792 sp:FEPC_ECOLI Escherichia coli K12 fepC 56.2 83.7 258 ferric enterobactin transport ATP- binding protein 562 4062 530626 529592 1035 sp:FEPG_ECOLI Escherichia coli K12 fepG 45.6 77.8 329 ferric enterobactin transport protein 563 4063 531782 530748 1035 sp:FEPD_ECOLI Escherichia coli K12 fepD 48.1 80.6 335 ferric enterobactin transport protein 564 4064 532008 532523 516 gp:CTACTAGEN_1 Thermoanaerobacterium 56.6 79.3 145 butyryl-CoA: acetate coenzyme A thermosaccharolyticum actA transferase 565 4065 533099 533401 303 sp:RS10_PLARO Planobispora rosea ATCC 84.2 99.0 101 30S ribosomal protein S10 53733 rpsJ 566 4066 533437 534090 654 sp:RL3_MYCBO Mycobacterium bovis BCG rplC 66.5 89.6 212 50S ribosomal protein L3 567 4067 534087 533401 687 568 4068 534090 534743 654 sp:RL4_MYCBO Mycobacterium bovis BCG rplD 71.2 90.1 212 50S ribosomal protein L4 569 4069 534746 535048 303 sp:RL23_MYCBO Mycobacterium bovis BCG rplW 74.0 90.6 96 50S ribosomal protein L23 570 4070 535072 534746 327 571 4071 535076 535915 840 sp:RL2_MYCLE Mycobacterium bovis BCG rplB 80.7 92.9 280 50S ribosomal protein L2 572 4072 535935 536210 276 sp:RS19_MYCTU Mycobacterium tuberculosis 87.0 98.9 92 30S ribosomal protein S19 H37Rv Rv0705 rpsS 573 4073 536183 535899 285 574 4074 536217 536576 360 sp:RL22_MYCTU Mycobacterium tuberculosis 74.3 91.7 109 50S ribosomal protein L22 H37Rv Rv0706 rplV 575 4075 536579 537322 744 sp:RS3_MYCBO Mycobacterium bovis BCG rpsC 77.4 91.2 239 30S ribosomal protein S3 576 4076 537328 537741 414 sp:RL16_MYCBO Mycobacterium bovis BCG rplP 69.3 88.3 137 50S ribosomal protein L16 577 4077 537744 537971 228 sp:RL29_MYCBO Mycobacterium bovis BCG rpmC 65.7 88.1 67 50S ribosomal protein L29 578 4078 537977 538252 276 sp:RS17_MYCBO Mycobacterium bovis BCG rpsQ 69.5 89.0 82 30S ribosomal protein S17 579 4079 538267 537974 294 580 4080 538698 538381 318 581 4081 539413 538718 696 582 4082 539741 540106 366 sp:RL14_MYCTU Mycobacterium tuberculosis 83.6 95.1 122 50S ribosomal protein L14 H37Rv Rv0714 rplN 583 4083 540112 540423 312 sp:RL24_MYCTU Mycobacterium tuberculosis 76.2 91.4 105 50S ribosomal protein L24 H37Rv Rv0715 rplX 584 4084 540426 540998 573 sp:RL5_MICLU Micrococcus luteus rplE 73.6 92.3 183 50S ribosomal protein L5 585 4085 541048 542079 1032 586 4086 542896 542090 807 sp:2DKG_CORSP Corynebacterium sp. 52.3 74.2 260 2,5-diketo-D-gluconic acid reductase 587 4087 543412 542921 492 588 4088 544329 543415 915 sp:FDHD_WOLSU Wolinella succinogenes fdhD 28.9 59.7 298 formate dehydrogenase chain D 589 4089 544670 544335 336 gp:SCGD3_29 Streptomyces coelicolor A3(2) 37.2 68.1 94 molybdopterin-guanine dinucleotide SCGD3.29c biosynthesis protein 590 4090 546889 544757 2133 sp:FDHF_ECOLI Escherichia coli fdfF 24.3 53.4 756 formate dehydrogenase H or alpha chain 591 4091 547329 548084 756 592 4092 548990 548187 804 593 4093 550651 548990 1662 sp:YC81_MYCTU Mycobacterium tuberculosis 26.9 52.6 624 ABC transporter ATP-binding protein H37Rv Rv1281c oppD 594 4094 551844 550699 1146 595 4095 552927 551854 1074 596 4096 554129 552948 1182 pir:E69424 Archaeoglobus fulgidus AF1398 24.7 50.4 405 hypothetical protein 597 4097 554919 554452 468 gp:AE001931_13 Deinococcus radiodurans 42.7 66.7 150 hypothetical protein DR0763 598 4098 555331 555726 396 pir:S29885 Micrococcus luteus 75.8 97.7 132 30S ribosomal protein S8 599 4099 555749 556282 534 pir:S29886 Micrococcus luteus 59.2 87.7 179 50S ribosomal protein L6 600 4100 556289 556690 402 sp:RL18_MICLU Micrococcus luteus rplR 67.3 90.9 110 50S ribosomal protein L18 601 4101 556734 557366 633 sp:RS5_MICLU Micrococcus luteus rpsE 67.8 88.3 171 30S ribosomal protein S5 602 4102 557373 557555 183 sp:RL30_ECOLI Escherichia coli K12 rpmJ 54.6 76.4 55 50S ribosomal protein L30 603 4103 557565 558008 444 sp:RL15_MICLU Micrococcus luteus rplO 66.4 87.4 143 50s ribosomal protein L15 604 4104 557588 556860 729 605 4105 558517 558197 321 prf.2204281A Streptomyces coelicolor msdA 46.9 68.8 128 methylmalonic acid semialdehyde dehydrogenase 606 4106 558969 558607 363 607 4107 559805 560260 456 GP:ABCARRA_2 Azospirillum brasilense carR 47.0 52.0 125 novel two-component regulatory system 608 4108 560634 559144 1491 prf.2516398E Rhodococcus rhodochrous 41.7 71.5 487 aldehyde dehydrogenase or betaine plasmid pRTL1 orf5 aldehyde dehydrogenase 609 4109 561368 560634 735 610 4110 562632 562937 306 611 4111 562633 561368 1266 prf:2411257B Sphingomonas sp. redA2 41.1 71.6 409 reductase 612 4112 562963 562646 318 prf:2313248B Rhodobacter capsulatus fdxE 47.7 66.4 107 2Fe2S ferredoxin 613 4113 563736 562993 744 gp:PPU24215_2 Pseudomonas putida cymB 35.8 70.8 257 p-cumic alcohol dehydrogenase 614 4114 563871 564083 213 PIR:H72754 Aeropyrum pernix K1 APE0029 50.0 56.0 50 hypothetical protein 615 4115 565471 563732 1740 pir:JC4176 Pyrococcus furiosus Vc1 DSM 22.9 45.0 629 phosphoenolpyruvate synthetase 3638 ppsA 616 4116 566759 565680 1080 pir:JC4176 Pyrococcus furiosus Vc1 DSM 38.6 66.7 378 phosphoenolpyruvate synthetase 3638 ppsA 617 4117 568088 566799 1290 prf:2104333G Rhodococcus erythropolis thcB 34.8 65.2 422 cytochrome P450 618 4118 569075 568272 804 prf:2512309A Erwinia carotovora carotovora 28.5 66.0 256 transcriptional repressor kdgR 619 4119 570774 571316 543 sp:KAD_MICLU Micrococcus luteus adk 48.9 81.0 184 adenylate kinase 620 4120 571367 570756 612 621 4121 571476 572267 792 sp:AMPM_BACSU Bacillus subtilis 168 map 43.1 74.7 253 methionine aminopeptidase 622 4122 572349 573176 828 623 4123 573407 573622 216 pir:F69644 Bacillus subtilis infA 77.0 86.0 72 translation initiation factor IF-1 624 4124 573816 574181 366 prf:2505353B Thermus thermophilus HB8 66.4 91.0 122 30S ribosomal protein S13 rps13 625 4125 574187 574588 402 sp:RS11_STRCO Streptomyces coelicolor A3(2) 81.3 93.3 134 30S ribosomal protein S11 SC6G4.06. rpsK 626 4126 574615 575217 603 prf:2211287F Mycobacterium tuberculosis 82.6 93.9 132 30S ribosomal protein S4 H37Rv RV3458C rpsD 627 4127 575338 576351 1014 sp:RPOA_BACSU Bacillus subtilis 168 rpoA 51.1 77.8 311 RNA polymerase alpha subunit 628 4128 575366 575211 156 629 4129 576410 576898 489 sp:RL17_ECOLI Escherichia coli K12 rplQ 51.6 77.1 122 50S ribosomal protein L17 630 4130 577057 577923 867 sp:TRUA_ECOLI Escherichia coli K12 truA 37.0 61.1 265 pseudouridylate synthase A 631 4131 578033 580429 2397 pir:G70695 Mycobacterium tuberculosis 24.8 51.2 786 hypothetical membrane protein H37Rv Rv3779 632 4132 580891 580436 456 633 4133 581221 580919 303 634 4134 581406 582662 1257 pir:A70836 Mycobacterium tuberculosis 27.4 53.8 485 hypothetical protein H37Rv Rv0283 635 4135 582684 584228 1545 sp:DIM_ARATH Arabidopsis thaliana CV DIM 22.8 50.9 505 cell elongation protein 636 4136 584268 585620 1353 sp:CFA_ECOLI Escherichia coli K12 cfa 30.7 56.0 423 cyclopropane-fatty-acyl-phospholipid synthase 637 4137 585823 586248 426 gp:SCL2_30 Streptomyces coelicolor A3(2) 28.0 59.0 100 hypothetical membrane protein SCL2.30c 638 4138 587757 586399 1359 sp:ELYA_BACAO Bacillus alcalophilus 31.3 58.0 273 high-alkaline serine proteinase 639 4139 589015 587645 1371 pir:T10930 Streptomyces coelicolor A3(2) 24.0 50.6 516 hypothetical membrane protein SC3C3.21 640 4140 589296 592862 3567 pir:E70977 Mycobacterium tuberculosis 65.0 38.4 1260 hypothetical membrane protein H37Rv Rv3447c 641 4141 590411 589590 822 642 4142 590560 589898 663 643 4143 592862 593761 900 644 4144 593935 594258 324 pir:C70977 Mycobacterium tuberculosis 31.1 69.9 103 hypothetical protein H37Rv Rv3445c 645 4145 594293 594580 288 prf:2111376A Mycobacterium tuberculosis 36.3 81.3 80 early secretory antigen target ESAT- 6 protein 646 4146 594939 595379 441 sp:RL13_STRCO Streptomyces coelicolor A3(2) 58.6 82.1 145 50S ribosomal protein L13 SC6G4.12. rplM 647 4147 595382 595927 546 sp:RS9_STRCO Streptomyces coelicolor A3(2) 49.2 72.4 181 30S ribosomal protein S9 SC6G4.13. rpsl 648 4148 596109 597449 1341 prf:2320260A Staphylococcus aureus 48.9 76.4 450 phosphoglucosamine mutase femR315 649 4149 597892 598194 303 650 4150 598194 599702 1509 pir:S75138 Synechocystis sp. PCC6803 29.3 45.6 318 hypothetical protein slr1753 651 4151 599350 598778 573 652 4152 599699 599932 234 653 4153 600876 600022 855 pir:S73000 Mycobacterium leprae 44.0 72.2 259 hypothetical protein B229_F1_20 654 4154 600971 602053 1083 sp:ALR_MYCTU Mycobacterium tuberculosis 41.6 68.5 368 alanine racemase H37Rv RV3423C alr 655 4155 602080 602574 495 sp:Y097_MYCTU Mycobacterium tuberculosis 48.7 78.6 154 hypothetical protein H37Rv Rv3422c 656 4156 602811 604409 1599 sp:YIDE_ECOLI Escherichia coli K12 yidE 28.9 66.2 550 hypothetical membrane protein 657 4157 604470 605708 1239 gp:PSJ00161_1 Propionibacterium shermanii pip 51.3 77.6 411 proline iminopeptidase 658 4158 605718 606392 675 sp:Y098_MYCTU Mycobacterium tuberculosis 52.2 75.4 207 hypothetical protein H37Rv Rv3421c 659 4159 606392 606898 507 sp:RIMI_ECOLI Escherichia coli K12 riml 30.3 59.9 132 ribosomal-protein-alanine N- acetyltransferase 660 4160 606905 607936 1032 sp:GCP_PASHA Pasteurella haemolytica 46.1 75.2 319 O-sialoglycoprotein endopeptidase SEROTYPE A1 gcp 661 4161 607958 609679 1722 sp:Y115_MYCTU Mycobacterium tuberculosis 38.4 59.4 571 hypothetical protein H37Rv Rv3433c 662 4162 609747 610175 429 663 4163 610268 609816 453 664 4164 610348 610644 297 sp:CH10_MYCTU Mycobacterium tuberculosis 76.0 94.0 100 heat shock protein groES H37Rv RV3418C mopB 665 4165 610659 612272 1614 sp:CH61_MYCLE Mycobacterium leprae 63.3 85.1 537 heat shock protein groEL B229_C3_248 groE1 666 4166 611200 610946 255 GP:MSGTCWPA_1 Mycobacterium tuberculosis 50.0 56.0 76 hypothetical protein 667 4167 612266 611109 1158 GP:MSGTCWPA_9 Mycobacterium tuberculosis 34.0 45.0 138 hypothetical protein 668 4168 612714 612418 297 gp:AF073300_1 Mycobacterium smegmatis 64.9 88.3 94 regulatory protein whiB3 669 4169 613156 613719 564 sp:Y09F_MYCTU Mycobacterium tuberculosis 55.2 81.6 174 RNA polymerase sigma factor H37Rv Rv3414c sigD 670 4170 613722 614747 1026 671 4171 615180 614803 378 sp:Y09H_MYCLE Mycobacterium leprae 41.4 69.8 116 hypothetical protein B1620_F3_131 672 4172 615336 616853 1518 gp:AB003154_1 Corynebacterium 80.8 93.9 504 IMP dehydrogenase ammoniagenes ATCC 6872 guaB 673 4173 616231 615605 627 PIR:F71456 Pyrococcus horikoshii PH0308 39.0 53.0 146 hypothetical protein 674 4174 616973 618094 1122 gp:AB003154_2 Corynebacterium 70.9 86.1 381 IMP dehydrogenase ammoniagenes ATCC 6872 675 4175 619013 618093 921 sp:YBIF_ECOLI Escherichia coli K12 ybiF 38.0 67.5 274 hypothetical membrane protein 676 4176 619086 619994 909 prf:1516239A Bacillus subtilis gltC 29.0 58.4 262 glutamate synthetase positive regulator 677 4177 620004 621572 1569 sp:GUAA_CORAM Corynebacterium 81.6 92.8 517 GMP synthetase ammoniagenes guaA 678 4178 620926 620264 663 679 4179 621717 622157 441 680 4180 622269 622457 189 681 4181 623635 622460 1176 gp:SCD63_22 Streptomyces coelicolor A3(2) 20.5 39.6 513 hypothetical membrane protein 682 4182 623800 624939 1140 gp:SC6E10_15 Streptomyces coelicolor A3(2) 26.8 48.7 411 two-component system sensor SC6E10.15c histidine kinase 683 4183 624985 625674 690 sp:DEGU_BACSU Bacillus subtilis 168 degU 33.5 65.1 218 transcriptional regulator or extracellular proteinase response regulator 684 4184 625677 626000 324 685 4185 626558 626070 489 686 4186 627539 626577 963 687 4187 627727 628551 825 pir:B70975 Mycobacterium tuberculosis 30.9 64.2 201 hypothetical protein H37Rv Rv3395c 688 4188 628551 630140 1590 pir:A70975 Mycobacterium tuberculosis 37.5 64.1 563 hypothetical protein H37Rv Rv3394c 689 4189 630810 630151 660 690 4190 630949 631809 861 gp:SC5B8_20 Streptomyces coelicolor A3(2) 33.8 62.9 275 hypothetical protein SC5B8.20c 691 4191 632684 631824 861 gp:AE001935_7 Deinococcus radiodurans 27.8 58.3 288 hypothetical membrane protein DR0809 692 4192 633079 632690 390 693 4193 633474 633079 396 gp:MMU92075_3 Mycobacterium marinum 36.8 67.4 95 hypothetical membrane protein 694 4194 635175 633532 1644 gp:AF139916_3 Brevibacterium linens ATCC 50.4 76.2 524 phytoene desaturase 9175 crtl 695 4195 636089 635178 912 gp:AF139916_2 Brevibacterium linens ATCC 42.0 71.2 288 phytoene synthase 9175 crtB 696 4196 638278 636089 2190 gp:SCF43A_29 Streptomyces coelicolor A3(2) 48.6 75.6 722 transmembrane transport protein SCF43A.29c 697 4197 639462 638317 1146 gp:AF139916_11 Brevibacterium linens crtE 32.7 63.8 367 geranylgeranyl pyrophosphate (GGPP) synthase 698 4198 639624 640208 585 gp:AF139916_14 Brevibacterium linens 38.3 68.1 188 transcriptional regulator (MarR family) 699 4199 640879 640232 648 sp:BLC_CITFR Citrobacter freundii blc OS60 blc 33.1 62.1 145 outer membrane lipoprotein 700 4200 641133 642557 1425 gp:AF139916_1 Brevibacterium linens 48.7 74.2 462 hypothetical protein 701 4201 643959 642556 1404 gp:AF139916_5 Brevibacterium linens ATCC 40.0 63.2 497 DNA photolyase 9175 cpd 1 702 4202 644026 644778 753 gp:AF155804_7 Streptococcus suis cps 1K 25.9 53.7 205 glycosyl transferase 703 4203 647590 645176 2415 gp:SCE25_30 Streptomyces coelicolor A3(2) 24.3 54.9 897 ABC transporter SCE25.30 704 4204 648309 647593 717 prf:2420410P Bacillus subtilis 168 yvrO 35.4 72.2 223 ABC transporter 705 4205 648467 648315 153 706 4206 649105 648440 666 prf:2320284D Helicobacter pylori abcD 35.9 75.2 206 ABC transporter 707 4207 649342 650187 846 708 4208 650193 649114 1080 sp:ABC_ECOLI Escherichia coli TAP90 abc 43.6 75.4 346 ABC transporter 709 4209 651288 650392 897 sp:HLPA_HAEIN Haemophilus influenzae 28.7 67.2 268 lipoprotein SEROTYPE B hlpA 710 4210 651601 654612 3012 prf:2517386A Thermus aquaticus dnaE 30.2 57.5 1101 DNA polymerase III 711 4211 654676 655122 447 gp:SCE126_11 Streptomyces coelicolor A3(2) 41.5 62.3 159 hypothetical protein SCE126.11 712 4212 655122 656534 1413 gp:SCE9_1 Streptomyces coelicolor A3(2) 26.1 56.0 468 hypothetical membrane protein SCE9.01 713 4213 655834 655097 738 714 4214 656547 657215 669 pir:C70884 Mycobacterium tuberculosis 50.3 76.4 203 transcriptional repressor H37Rv Rv2788 sirR 715 4215 658002 657205 798 gp:SCG8A_5 Streptomyces coelicolor A3(2) 34.9 61.7 264 hypothetical protein SCG8A.05c 716 4216 658005 658142 138 717 4217 658155 658928 774 pir:C69459 Archaeoglobus fulgidus AF1676 42.5 71.8 245 transcriptional regulator (Sir2 family) 718 4218 658933 659424 492 gp:SC5H1_34 Streptomyces coelicolor A3(2) 45.2 78.3 157 hypothetical protein SC5H1.34 719 4219 659543 660538 996 gp:CDU02617_1 Corynebacterium diphtheriae 31.1 62.2 357 iron-regulated lipoprotein precursor irp1 720 4220 661120 660650 471 pir:E70971 Mycobacterium tuberculosis 62.9 86.1 151 rRNA methylase H37Rv Rv3366 spoU 721 4221 661166 662017 852 pir:C70970 Mycobacterium tuberculosis 70.9 87.4 278 methylenetetrahydrofolate H37Rv Rv3356c folD dehydrogenase 722 4222 662120 662374 255 gp:MLCB1779_8 Mycobacterium leprae 31.3 76.3 80 hypothetical membrane protein MLCB1779.16c 723 4223 663761 662382 1380 gp:SC66T3_18 Streptomyces coelicolor A3(2) 34.0 63.2 489 hypothetical protein SC66T3.18c 724 4224 665088 664126 963 725 4225 666313 665183 1131 gp:AF052652_1 Corynebacterium glutamicum 99.5 99.5 379 homoserine O-acetyltransferase metA 726 4226 667770 666460 1311 prf:2317335A Leptospira meyeri metY 49.7 76.2 429 O-acetylhomoserine sulfhydrylase 727 4227 668264 670465 2202 sp:CSTA_ECOLI Escherichia coli K12 cstA 53.9 78.4 690 carbon starvation protein 728 4228 670053 669445 609 729 4229 670472 670672 201 sp:YJIX_ECOLI Escherichia coli K12 yjiX 40.0 66.0 50 hypothetical protein 730 4230 671653 671045 609 731 4231 671700 672653 954 pir:C70539 Mycobacterium tuberculosis 71.0 86.4 317 hypothetical protein H37Rv Rv1130 732 4232 672665 673576 912 prf:1902224A Streptomyces hygroscopicus 41.6 76.2 281 carboxy phosphoenolpyruvate mutase 733 4233 673608 674756 1149 sp:CISY_MYCSM Mycobacterium smegmatis 56.1 81.3 380 citrate synthase ATCC 607 gltA 734 4234 673639 672710 930 735 4235 674990 674799 192 sp:YNEC_ECOLI Escherichia coli K12 yneC 34.0 62.3 53 hypothetical protein 736 4236 675175 675846 672 737 4237 676122 675082 1041 sp:MDH_METFE Methanothermus fervidus V24S 37.6 67.5 338 L-malate dehydrogenase mdh 738 4238 676937 676218 720 prf.2514353L Bacillus stearothermophilus T-6 26.1 62.8 226 regulatory protein uxuR 739 4239 677748 677047 702 740 4240 681027 680131 897 sp:VIUB_VIBCH Vibrio cholerae OGAWA 395 25.4 54.2 284 vibriobactin utilization protein viuB 741 4241 681846 681040 807 gp:AF176902_3 Corynebacterium diphtheriae 55.4 85.1 269 ABC transporter ATP-binding protein irp 1D 742 4242 682904 681846 1059 gp:AF176902_2 Corynebacterium diphtheriae 56.3 86.4 339 ABC transporter irp 1C 743 4243 683866 682871 996 gp:AF176902_1 Corynebacterium diphtheriae 63.0 88.2 330 ABC transporter irp 1B 744 4244 684925 683876 1050 gp:CDU02617_1 Corynebacterium diphtheriae 53.1 82.3 356 iron-regulated lipoprotein precursor irp 1 745 4245 685109 686380 1272 prf:2202262A Streptomyces venezuelae cmlv 32.2 69.6 395 chloramphenicol resistance protein 746 4246 686435 687346 912 prf:2222220B Pseudomonas aeruginosa crc 30.4 58.1 303 catabolite repression control protein 747 4247 687351 688007 657 sp:YICG_HAEIN Haemophilus influenzae Rd 56.2 85.8 219 hypothetical protein HI1240 748 4248 688141 688335 195 749 4249 689890 688916 975 750 4250 690696 689917 780 gp:AF109162_3 Corynebacterium diphtheriae 45.1 73.8 244 ferrichrome ABC transporter hmuV 751 4251 691722 690706 1017 pir:S54438 Yersinia enterocolitica hemU 38.7 69.1 346 hemin permease 752 4252 691882 692916 1035 sp:SYW_ECOLI Escherichia coli K12 trpS 54.4 79.8 331 tryptophanyl-tRNA synthetase 753 4253 693028 694110 1083 sp:YHJD_ECOLI Escherichia coli K12 yhjD 37.1 72.3 278 hypothetical protein 754 4254 694172 695074 903 755 4255 696213 695077 1137 sp:DACD_SALTY Salmonella typhimurium LT2 30.9 57.5 301 penicillin-binding protein 6B dacD precursor 756 4256 697995 696769 1227 pir:F70842 Mycobacterium tuberculosis 34.1 70.7 417 hypothetical protein H37Rv Rv3311 757 4257 698922 698065 858 gp:SC6G10_8 Streptomyces coelicolor A3(2) 29.4 52.6 323 hypothetical protein SC6G10.08c 758 4258 699072 699266 195 759 4259 699272 698922 351 760 4260 699281 699913 633 sp:UPP_LACLA Lactococcus lactis upp 46.4 72.3 209 uracil phosphoribosyltransferase 761 4261 699998 700381 384 gp:SC1A2_11 Streptomyces coelicolor A3(2) 41.6 66.2 77 bacterial regulatory protein, lacl SC1A2.11 family 762 4262 702081 703262 1182 pir:H70841 Mycobacterium tuberculosis 51.4 80.5 385 N-acyl-L-amino acid amidohydrolase H37Rv Rv3305c amiA or peptidase 763 4263 702108 700384 1725 sp:MANB_MYCPI Mycoplasma pirum BER manB 22.1 53.8 561 phosphomannomutase 764 4264 703405 704811 1407 sp:DLDH_HALVO Halobacterium volcanii ATCC 31.6 65.0 468 dihydrolipoamide dehydrogenase 29605 lpd 765 4265 705211 708630 3420 prf:2415454A Corynebacterium glutamicum 100.0 100.0 1140 pyruvate carboxylase strain21253 pyc 766 4266 708839 709708 870 sp:YD24_MYCTU Mycobacterium tuberculosis 26.2 60.1 263 hypothetical protein H37Rv Rv1324 767 4267 709793 710278 486 gp:SCF11_30 Streptomyces coelicolor A3(2) 30.7 66.9 127 hypothetical protein SCF11.30 768 4268 711605 710520 1086 pir:B69760 Bacillus subtilis 168 yciC 44.6 69.0 381 hypothetical protein 769 4269 711724 712647 924 sp:TRXB_BACSU Bacillus subtilis IS58 trxB 24.6 59.3 305 thioredoxin reductase 770 4270 712738 714231 1494 sp:PRPD_SALTY Salmonella typhimurium LT2 24.0 49.5 521 PrpD protein for propionate prpD catabolism 771 4271 714258 715145 888 prf:1902224A Streptomyces hygroscopicus 42.5 74.5 278 carboxy phosphoenolpyruvate mutase 772 4272 714757 714380 378 PIR:E72779 Aeropyrum pernix K1 APE0223 39.0 47.0 96 hypothetical protein 773 4273 715102 716283 1182 sp:CISY_MYCSM Mycobacterium smegmatis 54.6 78.9 383 citrate synthase ATCC 607 gltA 774 4274 716660 716286 375 775 4275 718009 716687 1323 pir:B70539 Mycobacterium tuberculosis 40.8 72.6 456 hypothetical protein H37Rv Rv1129c 776 4276 718105 718350 246 777 4277 718658 720016 1359 778 4278 721449 720547 903 sp:THTR_CORGL Corynebacterium glutamicum 100.0 100.0 225 thiosulfate sulfurtransferase ATCC 13032 thtR 779 4279 721777 722841 1065 gp:CJ11168X1_62 Campylobacter jejuni Cj0069 61.1 79.8 352 hypothetical protein 780 4280 723338 722925 414 gp:MLCB4_16 Mycobacterium leprae 51.1 76.7 133 hypothetical protein MLCB4.27c 781 4281 723412 725559 2148 pir:G70539 Mycobacterium tuberculosis 35.1 63.4 718 hypothetical membrane protein H37Rv Rv1565c 782 4282 726462 725872 591 sp:YCEF_ECOLI Escherichia coli K12 yceF 31.8 66.2 192 hypothetical protein 783 4283 726715 726470 246 prf:2323363CF Mycobacterium leprae B1308- 33.3 69.8 63 hypothetical protein C3-211 784 4284 728352 726742 1611 gp:AB018531_2 Corynebacterium glutamicum 99.8 100.0 537 detergent sensitivity rescuer or AJ11060 dtsR2 carboxyl transferase 785 4285 730324 728696 1629 pir:JC4991 Corynebacterium glutamicum 99.6 100.0 543 detergent sensitivity rescuer or AJ11060 dtsR1 carboxyl transferase 786 4286 730436 731299 864 sp:BIRA_ECOLI Escherichia coli K12 birA 28.7 61.8 293 bifunctional protein (biotin synthesis repressor and biotin acetyl-CoA carboxylase ligase) 787 4287 731312 731797 486 pir:G70979 Mycobacterium tuberculosis 23.0 58.8 165 hypothetical membrane protein H37Rv Rv3278c 788 4288 731857 733017 1161 sp:PURK_CORAM Corynebacterium 69.0 83.8 394 5′-phosphoribosyl-5-amino-4- ammoniagenes ATCC 6872 imidasol carboxylase purK 789 4289 733072 734943 1872 sp:KUP_ECOLI Escherichia coli K12 kup 41.1 73.6 628 K+-uptake protein 790 4290 733797 733183 615 791 4291 734984 735340 357 792 4292 735402 735896 495 sp:PUR6_CORAM Corynebacterium 85.7 93.2 147 5′-phosphoribosyl-5-amiono-4- ammoniagenes ATCC 6872 imidasol carboxylase purE 793 4293 735899 736351 453 gp:APU33059_5 Actinosynnema pretiosum 36.2 60.5 152 hypothetical protein 794 4294 736413 737204 792 gp:SCF43A_36 Streptomyces coelicolor A3(2) 42.8 70.6 255 hypothetical protein SCF43A.36 795 4295 738529 737216 1314 sp:NTAA_CHEHE Chelatobacter heintzii ATCC 43.2 73.0 426 nitrilotriacetate monooxygenase 29600 ntaA 796 4296 740172 738673 1500 pir:A69426 Archaeoglobus fulgidus 23.4 52.5 303 transposase (ISA0963-5) 797 4297 741016 740228 789 sp:DHG2_BACME Bacillus megaterium IAM 1030 31.3 64.8 256 glucose 1-dehydrogenase gdhll 798 4298 741397 741765 369 pir:A72258 Thermotoga maritima MSB8 29.2 68.8 96 hypothetical membrane protein TM1408 799 4299 741854 742195 342 800 4300 742384 741818 567 sp:YWJB_BACSU Bacillus subtilis 168 ywjB 28.6 66.3 175 hypothetical protein 801 4301 742409 742828 420 gp:SCJ9A_21 Streptomyces coelicolor A3(2) 35.9 76.8 142 hypothetical protein SCJ9A.21 802 4302 743052 742831 222 803 4303 743900 743067 834 prf:2406355C Thermococcus litoralis malG 42.4 75.3 271 trehalose/maltose-binding protein 804 4304 744931 743900 1032 prf:2406355B Thermococcus litoralis malF 37.3 70.3 306 trehalose/maltose-binding protein 805 4305 745513 745046 468 806 4306 746893 745622 1272 prf:2406355A Thermococcus litoralis malE 30.9 62.4 417 trehalose/maltose-binding protein 807 4307 748020 748442 423 808 4308 748026 747031 996 prf:2308356A Streptomyces reticuli msiK 57.2 73.9 332 ABC transporter ATP-binding protein (ABC-type sugar transport protein) or cellobiose/maltose transport protein 809 4309 748446 748814 369 810 4310 753685 748886 4800 pir:B75633 Deinococcus radiodurans R1 25.1 49.9 1783 RNA helicase DRB0135 811 4311 757063 757434 372 812 4312 757395 753697 3699 813 4313 758262 757630 633 pir:E70978 Mycobacterium tuberculosis 31.7 59.2 240 hypothetical protein H37Rv Rv3268 814 4314 760796 758364 2433 pir:C71929 Helicobacter pylori J99 jhp0462 30.0 62.5 720 hypothetical protein 815 4315 762468 760906 1563 sp:UVRD_ECOLI Escherichia coli K12 uvrD 20.7 41.1 701 DNA helicase II 816 4316 762497 762853 357 817 4317 762730 763122 393 818 4318 762977 762582 396 819 4319 768191 767367 825 820 4320 769443 763237 6207 pir:T36671 Streptomyces coelicolor 22.4 45.8 2033 RNA helicase SCH5.13 821 4321 774142 769547 4596 pir:T08313 Halobacterium sp. NRC-1 24.4 53.2 698 hypothetical protein plasmid pNRC100 H1130 822 4322 777035 774150 2886 sp:HEPA_ECOLI Escherichia coli K12 hepA 23.1 48.6 873 RNA polymerase associated protein (ATP-dependent helicase) 823 4323 778711 777158 1554 pir:D70978 Mycobacterium tuberculosis 45.5 71.4 527 hypothetical protein H37Rv Rv3267 824 4324 779014 779910 897 gp:AF187550_1 Mycobacterium smegmatis 56.4 77.9 289 dTDP-Rha:a-D-GlcNAc- mc2155 wbbL diphosphoryl polyprenol, a-3-L- rhamnosyl transferase 825 4325 780128 781171 1044 sp:MPG1_YEAST Saccharomyces cerevisiae 29.8 66.9 353 mannose-1-phosphate YDL055C MPG1 guanylyltransferase 826 4326 781468 781875 408 gp:AF164439_1 Mycobacterium smegmatis 73.4 81.9 94 regulatory protein whmD 827 4327 782617 782162 456 pir:B70847 Mycobacterium tuberculosis 48.9 74.8 139 hypothetical protein H37Rv Rv3259 828 4328 782712 783101 390 gp:SCE34_11 Streptomyces coelicolor A3(2) 51.5 71.3 136 hypothetical protein SCE34.11c 829 4329 783184 784557 1374 sp:MANB_SALMO Salmonella montevideo M40 38.0 66.3 460 phosphomannomutase manB 830 4330 784635 785639 1005 pir:B70594 Mycobacterium tuberculosis 31.2 56.3 327 hypothetical protein H37Rv Rv3256c 831 4331 785643 786824 1182 sp:MANA_ECOLI Escherichia coli K12 manA 36.9 66.2 420 mannose-6-phosphate isomerase 832 4332 786896 787045 150 833 4333 787624 787983 360 834 4334 787733 787170 564 prf:1804279K Enterococcus faecalis plasmid 35.6 57.8 180 pheromone-responsive protein pCF10 prgC 835 4335 788196 788546 351 836 4336 788672 790093 1422 sp:SAHH_TRIVA Trichomonas vaginalis WAA38 59.0 83.0 476 S-adenosyl-L-homocysteine hydrolase 837 4337 789426 788719 708 838 4338 789721 789002 720 839 4339 790096 790704 609 sp:KTHY_ARCFU Archaeoglobus fulgidus VC-16 25.8 56.0 209 thymidylate kinase AF0061 840 4340 790732 791409 678 prf:2214304A Mycobacterium tuberculosis 73.7 90.6 224 two-component system response H37Rv Rv3246c mtrA regulator 841 4341 791421 790738 684 842 4342 791512 793008 1497 prf:2214304B Mycobacterium tuberculosis 53.1 78.9 484 two-component system sensor H37Rv Rv3245c mtrB histidine kinase 843 4343 793008 794711 1704 pir:F70592 Mycobacterium tuberculosis 29.6 65.6 595 lipoprotein H37Rv Rv3244c lpqB 844 4344 794714 795301 588 pir:D70592 Mycobacterium tuberculosis 38.0 72.8 213 hypothetical protein H37Rv Rv3242c 845 4345 795447 795292 156 846 4346 795448 796110 663 sp:RR3O_SPIOL Spinacia oleracea CV rps22 34.5 61.6 203 30S ribosomal protein or chloroplast precursor 847 4347 796250 798784 2535 gsp:R74093 Brevibacterium flavum 99.1 99.6 845 preprotein translocase SecA subunit (Corynebacterium glutamicum) MJ-233 secA 848 4348 799020 799691 672 849 4349 799697 800200 504 pir:A70591 Mycobacterium tuberculosis 47.1 78.8 170 hypothetical protein H37Rv Rv3231c 850 4350 801194 800208 987 pir:F70590 Mycobacterium tuberculosis 64.6 82.9 322 hypothetical protein H37Rv Rv3228 851 4351 802602 801190 1413 gp:AF114233_1 Corynebacterium glutamicum 99.0 99.0 461 5-enolpyruvylshikimate 3-phosphate ASO19 aroA synthase 852 4352 802649 803128 480 pir:D70590 Mycobacterium tuberculosis 38.3 63.9 180 hypothetical protein H37Rv Rv3226c 853 4353 802687 802565 123 GP:AF114233_1 Corynebacterium glutamicum 100.0 100.0 23 5-enolpyruvylshikimate 3-phosphate synthase 854 4354 804240 803131 1110 pir:G70506 Mycobacterium tuberculosis 21.6 42.4 380 hypothetical protein H37Rv Rv0336 855 4355 804408 805025 618 prf.2515333D Mycobacterium tuberculosis 61.2 87.2 188 RNA polymerase sigma factor sigH 856 4356 805792 805535 258 pir:D70596 Mycobacterium tuberculosis 78.6 96.4 84 regulatory protein H37Rv Rv3219 whiB1 857 4357 806318 806737 420 pir:B70596 Mycobacterium tuberculosis 33.3 65.1 129 hypothetical protein H37Rv Rv3217c 858 4358 807939 806740 1200 pir:E70595 Mycobacterium tuberculosis 29.6 62.2 415 hypothetical protein H37Rv Rv3212 859 4359 809217 807946 1272 sp:DEAD_KLEPN Klebsiella pneumoniae CG43 37.3 64.0 458 DEAD box ATP-dependent RNA deaD helicase 860 4360 809286 809510 225 861 4361 809549 810394 846 pir:H70594 Mycobacterium tuberculosis 46.4 69.8 291 hypothetical protein H37Rv Rv3207c 862 4362 810405 811163 759 pir:F70594 Mycobacterium tuberculosis 37.0 65.9 249 hypothetical protein H37Rv Rv3205c 863 4363 811170 814217 3048 pir:G70951 Mycobacterium tuberculosis 23.9 48.9 1155 ATP-dependent DNA helicase H37Rv Rv3201c 864 4364 812165 811386 780 865 4365 814204 817422 3219 pir:G70951 Mycobacterium tuberculosis 41.4 65.7 1126 ATP-dependent DNA helicase H37Rv Rv3201c 866 4366 815541 814210 1332 867 4367 817519 818523 1005 sp:Y13B_METJA Methanococcus jannaschii JAL- 26.2 64.2 302 potassium channel 1 MJ0138.1 868 4368 818523 819236 714 pir:E70951 Mycobacterium tuberculosis 30.4 58.3 230 hypothetical protein H37Rv Rv3199c 869 4369 819254 821287 2034 sp:UVRD_ECOLI Escherichia coli K12 uvrD 32.6 58.8 660 DNA helicase II 870 4370 822079 822669 591 871 4371 822105 821290 816 pir:B70951 Mycobacterium tuberculosis 26.8 49.3 280 hypothetical protein H37Rv Rv3196 872 4372 822789 823391 603 873 4373 824125 822680 1446 pir:A70951 Mycobacterium tuberculosis 42.8 76.4 474 hypothetical protein H37Rv Rv3195 874 4374 824190 825239 1050 pir:H70950 Mycobacterium tuberculosis 43.4 74.9 350 hypothetical protein H37Rv Rv3194 875 4375 825916 825242 675 876 4376 826517 825996 522 877 4377 826616 829570 2955 pir:G70950 Mycobacterium tuberculosis 47.2 73.5 1023 hypothetical protein H37Rv Rv3193c 878 4378 830985 829627 1359 gp:AE001938_5 Deinococcus radiodurans 34.3 57.7 463 regulatory protein DR0840 879 4379 831021 831971 951 sp:ER1_HEVBR Hevea brasiliensis laticifer er1 67.4 89.0 301 ethylene-inducible protein 880 4380 831922 831578 345 PIR:F72782 Aeropyrum pernix K1 APE0247 49.0 53.0 81 hypothetical protein 881 4381 831971 832570 600 sp:YAAE_BACSU Bacillus subtilis 168 yaaE 40.8 73.6 201 hypothetical protein 882 4382 833157 832795 363 883 4383 833572 834633 1062 pir:TRYXB4 Lysobacter enzymogenes ATCC 26.7 44.4 408 alpha-lytic proteinase precursor 29487 884 4384 834888 835388 501 885 4385 835253 835837 585 pir:S03722 Neurospora intermedia LaBelle- 25.0 51.4 208 DNA-directed DNA polymerase 1b mitochondrion plasmid 886 4386 837312 838892 1581 sp:CSP1_CORGL Corynebacterium glutamicum 27.0 51.5 363 major secreted protein PS1 protein (Brevibacterium flavum) ATCC precursor 17965 csp1 887 4387 838925 839353 429 888 4388 839630 840139 510 889 4389 840431 840210 222 890 4390 840745 840437 309 891 4391 842296 841517 780 prf:2207273H Streptomyces alboniger pur3 51.8 74.9 255 monophosphatase 892 4392 843124 842306 819 gp:U70376_9 Streptomyces flavopersicus 33.7 59.3 243 myo-inositol monophosphatase spcA 893 4393 843257 844360 1104 sp:RF2_STRCO Streptomyces coelicolor A3(2) 68.0 88.6 359 peptide chain release factor 2 prfB 894 4394 844495 845181 687 pir:E70919 Mycobacterium tuberculosis 70.4 91.2 226 cell division ATP-binding protein H37Rv Rv3102c ftsE 895 4395 845105 844842 264 PIR:G72510 Aeropyrum pernix K1 APE2061 43.0 54.0 72 hypothetical protein 896 4396 845198 846097 900 pir:D70919 Mycobacterium tuberculosis 40.5 74.8 301 cell division protein H37Rv Rv3101c ftsX 897 4397 846137 846628 492 sp:SMPB_ECOLI Escherichia coli K12 smpB 43.5 75.9 145 small protein B (SSRA-binding protein) 898 4398 846632 846982 351 sp:YEAO_ECOLI Escherichia coli K12 yeaO 44.0 73.3 116 hypothetical protein 899 4399 846805 846269 537 900 4400 847727 848026 300 901 4401 848122 847718 405 902 4402 849323 848499 825 sp:VIUB_VIBCH Vibrio cholerae OGAWA 395 26.8 52.9 272 vibriobactin utilization protein viuB 903 4403 850243 849326 918 prf:2510361A Staphylococcus aureus sirA 29.5 58.3 319 Fe-regulated protein 904 4404 850999 850412 588 gp:MLCB1243_5 Mycobacterium leprae 36.1 71.2 191 hypothetical membrane protein MLCB1243.07 905 4405 851351 852364 1014 sp:FATB_VIBAN Vibrio anguillarum 775 fatB 27.7 61.5 325 ferric anguibactin-binding protein precursor 906 4406 852618 853616 999 pir:B69763 Bacillus subtilis 168 yclN 39.3 80.8 313 ferrichrome ABC transporter (permease) 907 4407 853783 854724 942 pir:C69763 Bacillus subtilis 168 yclO 35.6 76.0 312 ferrichrome ABC transporter (permease) 908 4408 854724 855476 753 pir:D69763 Bacillus subtilis 168 yclP 48.4 82.0 250 ferrichrome ABC transporter (ATP- binding protein) 909 4409 860224 860078 147 PIR:F81737 Chlamydia muridarum Nigg 66.0 72.0 48 hypothetical protein TC0129 910 4410 860745 860473 273 GSP:Y35814 Chlamydia pneumoniae 61.0 66.0 84 hypothetical protein 911 4411 861544 862752 1209 pir:S66270 Rattus norvegicus (Rat) 33.5 64.9 442 kynurenine aminotransferase/glutamine transaminase K 912 4412 863391 862753 639 913 4413 865066 863396 1671 sp:RA25_YEAST Saccharomyces cerevisiae 30.7 62.3 613 DNA repair helicase S288C YIL143C RAD25 914 4414 867317 865119 2199 pir:F70815 Mycobacterium tuberculosis 36.1 65.2 764 hypothetical protein H37Rv Rv0862c 915 4415 867353 867571 219 pir:G70815 Mycobacterium tuberculosis 44.0 62.0 57 hypothetical protein H37Rv Rv0863 916 4416 867788 868630 843 917 4417 868399 867803 597 prf:2420502A Micrococcus luteus rpf 39.4 64.7 198 resuscitation-promoting factor 918 4418 868938 869318 381 prf:2320271A Lactococcus lactis cspB 42.6 75.4 61 cold shock protein 919 4419 869903 869379 525 gp:MLCB57_11 Mycobacterium leprae 28.3 58.5 159 hypothetical protein MLCB57.27c 920 4420 870691 869918 774 gp:AE001874_1 Deinococcus radiodurans 41.8 67.8 273 glutamine cyclotransferase DR0112 921 4421 871419 870721 699 922 4422 871523 871660 138 923 4423 871738 873210 1473 gp:SC6C5_9 Streptomyces coelicolor A3(2) 43.6 79.3 477 permease SC6C5.09 924 4424 872927 872016 912 925 4425 873213 874040 828 sp:TSNR_STRAZ Streptomyces azureus tsnR 27.9 51.7 319 rRNA(adenosine-2′-O-)- methyltransferase 926 4426 874944 874069 876 927 4427 875883 874951 933 sp:YZ11_MYCTU Mycobacterium tuberculosis 32.6 55.1 316 hypothetical protein H37Rv Rv0883c 928 4428 877112 875985 1128 pir:S71439 Bacillus circulans ATCC 21783 21.9 52.9 374 phosphoserine transaminase 929 4429 881114 879642 1473 sp:ACCD_ECOLI Escherichia coli K12 accD 36.0 69.5 236 acetyl-coenzyme A carboxylase carboxy transferase subunit beta 930 4430 881647 881985 339 gp:SCI8_8 Streptomyces coelicolor A3(2) 51.5 80.6 103 hypothetical protein SCI8.08c 931 4431 881995 883647 1653 pir:JC2382 Pseudomonas fluorescens 26.4 58.1 549 sodium/proline symporter 932 4432 883726 884541 816 933 4433 885388 884549 840 pir:A70657 Mycobacterium tuberculosis 49.0 77.4 243 hypothetical protein H37Rv Rv2525c 934 4434 885672 894578 8907 pir:S55505 Corynebacterium 63.1 83.4 3026 fatty-acid synthase ammoniagenes fas 935 4435 894703 895191 489 936 4436 895408 895593 186 937 4437 896642 895596 1047 prf:2317335B Leptospira meyeri metY metX 29.0 59.7 335 homoserine O-acetyltransferase 938 4438 897144 896719 426 939 4439 897423 897689 267 940 4440 897963 897727 237 gp:AE002044_8 Deinococcus radiodurans 43.6 72.6 62 glutaredoxin DR2085 941 4441 898434 897979 456 prf:2408256A Mycobacterium avium folA 38.0 62.0 17.1 dihydrofolate reductase 942 4442 899231 898434 798 sp:TYSY_ECOLI Escherichia coli K12 thyA 64.8 88.9 261 thymidylate synthase 943 4443 900008 899253 756 sp:CYSQ_ECOLI Escherichia coli K12 cysQ 32.2 56.4 202 ammonium transporter 944 4444 900043 904602 4560 gp:SC7C7_16 Streptomyces coelicolor A3(2) 47.4 68.1 1715 ATP dependent DNA helicase SC7C7.16c 945 4445 904615 905382 768 sp:FPG_SYNEN Synechococcus elongatus 29.2 51.0 298 formamidopyrimidine-DNA naegeli mutM glycosidase 946 4446 905389 905796 408 pir:F70816 Mycobacterium tuberculosis 55.5 86.7 128 hypothetical protein H37Rv Rv0870c 947 4447 906391 905792 600 sp:APL_LACLA Lactococcus lactis MG1363 apl 38.8 71.9 196 alkaline phosphatase 948 4448 907731 906559 1173 pir:T36776 Streptomyces coelicolor A3(2) 33.8 67.0 403 integral membrane transporter SCI28.06c 949 4449 908612 909328 717 950 4450 909378 907759 1620 pir:NUEC Escherichia coli JM101 pgi 52.4 77.0 557 glucose-6-phosphate isomease 951 4451 910696 909521 1176 pir:G70506 Mycobacterium tuberculosis 24.6 52.3 195 hypothetical protein H37Rv Rv0336 952 4452 910843 911223 381 953 4453 911163 910855 309 sp:YT26_MYCTU Mycobacterium tuberculosis 59.0 85.9 78 hypothetical protein H37Rv Rv0948c 954 4454 911226 913514 2289 sp:PCRA_BACST Bacillus stearothermophilus 46.1 73.1 763 ATP-dependent helicase NCA 1503 pcrA 955 4455 915699 913477 2223 gp:SCE25_30 Streptomyces coelicolor A3(2) 21.8 48.6 885 ABC transporter SCE25.30 956 4456 916364 915699 666 prf:2420410P Bacillus subtilis 168 yvrO 43.8 71.4 217 ABC transporter 957 4457 916874 916368 507 958 4458 917680 916970 711 pir:D70716 Mycobacterium tuberculosis 43.6 73.3 236 peptidase H37Rv Rv0950c 959 4459 917928 919352 1425 sp:YT19_MYCTU Mycobacterium tuberculosis 31.1 60.8 434 hypothetical protein H37Rv Rv0955 960 4460 918054 917827 228 961 4461 919330 919956 627 gp:AB003159_2 Corynebacterium 64.6 86.2 189 5′-phosphoribosylglycinamide ammoniagenes purN formyltransferase 962 4462 919967 921526 1560 gp:AB003159_3 Corynebacterium 74.5 87.8 525 5′-phosphoribosyl-5-aminoimidazole- ammoniagenes purH 4-carboxamide formyltransferase 963 4463 921594 922412 819 gp:CGL133719_3 Corynebacterium glutamicum 100.0 100.0 217 citrate lyase (subunit) ATCC 13032 citE 964 4464 923061 922396 666 gp:CGL133719_2 Corynebacterium glutamicum 100.0 100.0 222 repressor of the high-affinity (methyl) ATCC 13032 amtR ammonium uptake system 965 4465 923464 923138 327 gp:CGL133719_1 Corynebacterium glutamicum 100.0 100.0 109 hypothetical protein ATCC 13032 yjcC 966 4466 923661 923981 321 967 4467 924407 924159 249 sp:RR18_CYAPA Cyanophora paradoxa rps 18 52.2 76.1 67 30S ribosomal protein S18 968 4468 924727 924425 303 sp:RS14_ECOLI Escherichia coli K12 rpsN 54.0 80.0 100 30S ribosomal protein S14 969 4469 924895 924734 162 sp:RL33_ECOLI Escherichia coli K12 rpmG 55.1 83.7 49 50S ribosomal protein L33 970 4470 925134 924901 234 pir:R5EC28 Escherichia coli K12 rpmB 52.0 81.8 77 50S ribosomal protein L28 971 4471 926935 925325 1611 pir:B70033 Bacillus subtilis 168 yvdB 34.4 71.1 529 transporter (sulfate transporter) 972 4472 927242 926931 312 prf:2420312A Staphylococcus aureus zntR 37.5 77.5 80 Zn/Co transport repressor 973 4473 927474 927737 264 sp:RL31_HAEDU Haemophilus ducreyi rpmE 37.2 65.4 78 50S ribosomal protein L31 974 4474 927752 927922 171 gp:SC51A_14 Streptomyces coelicolor A3(2) 60.0 78.2 55 50S ribosomal protein L32 SCF51A 14 975 4475 927785 927339 447 976 4476 928117 928812 696 sp:COPR_PSESM Pseudomonas syringae copR 48.0 73.6 227 copper-inducible-two-component regulator 977 4477 928884 930248 1365 sp:BAES_ECOLI Escherichia coli K12 baeS 24.4 60.1 484 two-component system sensor 978 4478 930410 931648 1239 pir:S45229 Escherichia coli K12 htrA 33.3 59.9 406 proteinase DO precursor 979 4479 931706 932290 585 sp:CNXI_ARATH Arabidopsis thaliana CV cnxl 27.7 54.3 188 molybdopterin biosynthesis cnx1 protein (molybdenum cofactor biosynthesis enzyme cnx1) 980 4480 932290 932487 198 981 4481 932974 932570 405 sp:MSCL_MYCTU Mycobacterium tuberculosis 50.4 77.1 131 large-conductance H37Rv Rv0985c mscL mechanosensitive channel 982 4482 933710 933060 651 pir:A70601 Mycobacterium tuberculosis 28.6 60.0 210 hypothetical protein H37Rv Rv0990 983 4483 934302 933733 570 pir:JC4389 Homo sapiens MTHFS 25.1 59.7 191 5-formyltetrahydrofolate cyclo-ligase 984 4484 934423 935319 897 pir:JC4985 Xanthomonas campestris 42.2 68.9 296 UTP--glucose-1-phosphate uridylyltransferase 985 4485 935351 936607 1257 prf:2403296B Arthrobacter nicotinovorans 31.8 62.6 390 molybdopterin biosynthesis protein moeA 986 4486 936615 937274 660 sp:RIMJ_ECOLI Escherichia coli K12 rimJ 29.0 54.9 193 ribosomal-protein-alanine N- acetyltransferase 987 4487 937382 938401 1020 pir:G70601 Mycobacterium tuberculosis 30.3 54.8 367 hypothetical membrane protein H37Rv Rv0996 988 4488 938427 939626 1200 sp:CYNX_ECOLI Escherichia coli K12 cynX 26.6 62.4 380 cyanate transport protein 989 4489 939217 937799 1419 990 4490 939686 940090 405 sp:YG02_HAEIN Haemophilus influenzae Rd 32.1 60.6 137 hypothetical membrane protein HI1602 991 4491 940041 940754 714 sp:Y05C_MYCTU Mycobacterium tuberculosis 25.3 59.6 225 hypothetical membrane protein H37Rv Rv0093c 992 4492 940759 941925 1167 sp:CDAS_BACSH Bacillus sphaericus E-244 26.8 53.6 444 cyclomaltodextrinase CDase 993 4493 943940 942381 1560 pir:E70602 Mycobacterium tuberculosis 43.0 75.2 488 hypothetical membrane protein H37Rv 994 4494 944009 944833 825 sp:Y19J_MYCTU Mycobacterium tuberculosis 54.0 78.3 272 hypothetical protein H37Rv Rv1003 995 4495 946840 948669 1830 sp:SYM_METTH Methanobacterium 33.8 66.7 615 methionyl-tRNA synthetase thermoautotrophicum Delta H MTH587 metG 996 4496 948791 950839 2049 prf:1306383A Escherichia coli recQ 26.2 49.0 741 ATP-dependent DNA helicase 997 4497 951460 950828 633 pir:B69206 Methanobacterium 27.6 53.3 210 hypothetical protein thermoautotrophicum Delta H MTH796 998 4498 952991 951834 1158 sp:YXAG_BACSU Bacillus subtilis 168 yxaG 30.0 59.0 363 hypothetical protein 999 4499 953573 953043 531 1000 4500 953973 954266 294 gp:AF029727_1 Enterococcus faecium 33.0 59.6 94 transposase 1001 4501 954277 954753 477 pir:TQEC13 Escherichia coli K12 41.7 67.6 139 transposase 1002 4502 954941 955354 414 gp:AF052055_1 Brevibacterium linens tnpA 73.2 88.4 112 transposase subunit 1003 4503 955911 956774 864 1004 4504 957398 955686 1713 prf.2014253AE Escherichia coli dld 46.4 75.6 565 D-lactate dehydrogenase 1005 4505 958683 957844 840 sp:MTK1_KLEPN Kiebsielia pneumoniae 0K8 30.8 62.8 231 site-specific DNA-methyltransferase kpnlM 1006 4506 959403 959185 219 1007 4507 960081 960374 294 gp:AF029727_1 Enterococcus faecium 33.0 59.6 94 transposase 1008 4508 960385 960861 477 pir:TQEC13 Escherichia coli K12 41.7 67.6 139 transposase 1009 4509 961297 961653 357 sp:YJ94_MYCTU Mycobacterium tuberculosis 62.6 84.6 91 transcriptional regulator H37Rv Rv1994c 1010 4510 961629 962249 621 prf.2514367A Staphylococcus aureus cadD 31.7 66.8 205 cadmium resistance protein 1011 4511 961662 961321 342 1012 4512 962809 963639 831 pir:C70603 Mycobacterium tuberculosis 46.4 70.7 263 hypothetical protein H37Rv Rv1008 1013 4513 963864 964934 1071 pir:D70603 Mycobacterium tuberculosis 34.8 63.5 362 hypothetical protein H37Rv Rv1009 rpf 1014 4514 964974 965852 879 sp:KSGA_ECOLI Escherichia coli K12 ksgA 34.3 65.3 265 dimethyladenosine transferase 1015 4515 965852 966784 933 pir:F70603 Mycobacterium tuberculosis 42.5 67.0 315 isopentenyl monophosphate kinase H37Rv Rv1011 1016 4516 966591 965950 642 1017 4517 966828 968660 1833 pir:S47441 Saccharopolyspora erythraea 65.5 85.8 478 ABC transporter ertX 1018 4518 968667 969458 792 sp:PDXK_ECOLI Escherichia coli K12 pdxK 40.1 67.4 242 pyridoxine kinase 1019 4519 969940 969461 480 sp:YX05_MYCTU Mycobacterium tuberculosis 27.0 58.5 159 hypothetical protein H37Rv Rv2874 1020 4520 970029 970349 321 gp:SCF1_2 Streptomyces coelicolor A3(2) 45.4 78.7 108 hypothetical protein SCF1.02 1021 4521 970418 970738 321 gp:SCF1_2 Streptomyces coelicolor A3(2) 35.5 69.2 107 hypothetical protein SCF1.02 1022 4522 970864 971823 960 gp:SCJ1_15 Streptomyces coelicolor A3(2) 64.8 88.1 261 regulator SCJ1.15 1023 4523 973035 972244 792 sp:YXEH_BACSU Bacillus subtilis 168 yxeH 27.2 59.1 276 hypothetical protein 1024 4524 973139 974155 1017 pir:E70893 Mycobacterium tuberculosis 35.6 70.9 337 enoyl-CoA hydratase H37Rv echA9 1025 4525 973957 973304 654 1026 4526 974186 974962 777 1027 4527 976176 974965 1212 1028 4528 976349 977734 1386 sp:CSP1_CORGL Corynebacterium glutamicum 27.7 56.8 440 major secreted protein PS1 protein (Brevibacterium flavum) ATCC precursor 17965 csp1 1029 4529 978378 977800 579 gp:SCF56_6 Streptomyces coelicolor A3(2) 44.0 70.0 100 transcriptional regulator (tetR SCF56.06 family) 1030 4530 980740 978368 2373 gp:SCE87_17 Streptomyces coelicolor A3(2) 42.6 70.0 802 membrane transport protein SCE87.17c 1031 4531 980993 981490 498 sp:MENG_HAEIN Haemkophilus influenzae Rd 38.2 75.8 157 S-adenosylmethionine: 2- HI0508 menG demethylmenaquinone methyltransferase 1032 4532 981622 982287 666 1033 4533 982674 982294 381 gp:NMA6Z2491_214 Neisseria meningitidis NMA 1953 29.8 63.6 121 hypothetical protein 1034 4534 983100 984650 1551 pir:A70539 Mycobacterium tuberculosis 24.9 48.3 482 hypothetical protein H37Rv Rv1128c 1035 4535 984910 985845 936 1036 4536 986510 984864 1647 pir:I59305 Escherichia coli K12 prfC 39.2 68.0 546 peptide-chain-release factor 3 1037 4537 986739 988007 1269 prf.2406311A Methylophilus methylotrophus 42.8 72.8 404 amide-urea transport protein fmdD 1038 4538 988023 988904 882 prf:2406311B Methylophilus methylotrophus 40.8 61.0 77 amide-urea transport protein fmdE 1039 4539 988904 989980 1077 prf:2406311C Methylophilus methylotrophus 34.6 68.0 234 amide-urea transport protein fmdF 1040 4540 989980 990705 726 sp:BRAF_PSEAE Pseudomonas aeruginosa PAO 37.9 70.0 253 high-affinity branched-chain amino braF acid transport ATP-binding protein 1041 4541 990716 991414 699 sp:BRAG_PSEAE Pseudomonas aeruginosa PAO 35.2 69.1 236 high-affinity branched-chain amino braG acid transport ATP-binding protein 1042 4542 992028 991417 612 sp:PTH_ECOLI Escherichia coli K12 pth 39.0 70.6 187 peptidyl-tRNA hydrolase 1043 4543 992058 993080 1023 sp:2NPD_WILMR Williopsis mrakii IFO 0895 25.2 54.0 361 2-nitropropane dioxygenase 1044 4544 993549 994613 1065 sp:G3P_ZYMMO Streptomyces roseofulvus gap 39.5 72.8 342 glyceraldehyde-3-phosphate dehydrogenase 1045 4545 994474 994106 369 GSP:Y75094 Neisseria meningitidis 54.0 61.0 51 polypeptides predicted to be useful antigens for vaccines and diagnostics 1046 4546 995375 994845 531 sp:PTH_ECOLI Escherichia coli K12 pth 38.5 63.2 174 peptidyl-tRNA hydrolase 1047 4547 996126 995527 600 pir:B70622 Mycobacterium tuberculosis 47.0 65.0 194 50S ribosomal protein L25 H37Rv rplY 1048 4548 996402 996830 429 sp:LGUL_SALTY Salmonella typhimurium D21 28.7 54.6 143 lactoylglutathione lyase gloA 1049 4549 997456 996833 624 prf:2516401BW Bacillus cereus ATCC 10987 38.9 62.5 208 DNA alkylation repair enzyme alkD 1050 4550 998440 997466 975 sp:KPRS_BACCL Bacillus subtilis prs 44.0 79.1 316 ribose-phosphate pyrophosphokinase 1051 4551 999909 998455 1455 pir:S66080 Bacillus subtilis gcaD 42.0 71.9 452 UDP-N-acetylglucosamine pyrophosphorylase 1052 4552 1001242 1000016 1227 1053 4553 1001332 1002864 1533 sp:SUFI_ECOLI Escherichia coli K12 sufl 30.8 61.7 506 sufl protein precursor 1054 4554 1003013 1003930 918 sp:NODI_RHIS3 Rhizobium sp. N33 nodl 35.8 64.8 310 nodulation ATP-binding protein I 1055 4555 1003953 1004783 831 pir:JN0850 Streptomyces lividans ORF2 30.2 63.2 272 hypothetical membrane protein 1056 4556 1004829 1006085 1257 sp:UHPB_ECOLI Escherichia coli K12 uhpB 24.6 48.4 459 two-component system sensor histidine kinase 1057 4557 1006089 1006697 609 prf.2107255A Streptomyces peucetius dnrN 36.6 67.3 202 two component transcriptional regulator (luxR family) 1058 4558 1006937 1006734 204 1059 4559 1006998 1008152 1155 gp:SCF15_7 Streptomyces coelicolor A3(2) 31.5 64.5 349 hypothetical membrane protein SCF15.07 1060 4560 1008622 1010061 1440 pir:S65587 Streptomyces glaucescens strV 28.6 57.0 535 ABC transporter 1061 4561 1008686 1008534 153 1062 4562 1010057 1011790 1734 pir:T14180 Mycobacterium smegmatis exiT 44.0 74.0 573 ABC transporter 1063 4563 1013761 1011797 1965 sp:GGT_ECOLI Escherichia coli K12 ggt 32.4 58.6 666 gamma-glutamyltranspeptidase precursor 1064 4564 1014016 1014264 249 1065 4565 1014861 1014343 519 1066 4566 1014925 1015116 192 1067 4567 1015652 1016560 909 1068 4568 1015692 1015450 243 GPU:AF164956_23 Corynebacterium glutamicum 64.0 72.0 37 transposase protein fragment TnpNC 1069 4569 1015852 1015145 708 gp:AF121000_8 Corynebacterium glutamicum 99.6 100.0 236 transposase (IS1628 TnpB) 22243 R-plasmid pAG1 tnpB 1070 4570 1016557 1017018 462 1071 4571 1017870 1017274 597 1072 4572 1018082 1018393 312 1073 4573 1018416 1019066 651 sp:TETC_ECOLI Escherichia coli tetR 23.0 59.6 183 transcriptional regulator (TetR- family) 1074 4574 1019090 1022716 3627 sp:MFD_ECOLI Escherichia coli mfd 36.2 65.1 1217 transcription/repair-coupling protein 1075 4575 1020613 1019390 1224 1076 4576 1021305 1021078 228 GSP:Y75301 Neisseria gonorrhoeae 48.0 69.0 76 Neisserial polypeptides predicted to be useful antigens for vaccines and diagnostics 1077 4577 1024666 1022699 1968 sp:MDLB_ECOLI Escherichia coli mdlB 31.3 62.7 632 multidrug resistance-like ATP- binding protein, ABC-type transport protein 1078 4578 1026396 1024666 1731 sp:YC73_MYCTU Mycobacterium tuberculosis 50.2 81.9 574 ABC transporter H37Rv Rv1273c 1079 4579 1028886 1026505 2382 sp:YLI3_CORGL Corynebacterium glutamicum 100.0 100.0 368 hypothetical membrane protein ATCC 13032 orf3 1080 4580 1031885 1032181 297 1081 4581 1032196 1032780 585 sp.YABN_BACSU Bacillus subtilis yabN 33.4 57.4 183 hypothetical protein 1082 4582 1033185 1032760 426 1083 4583 1033646 1033269 378 1084 4584 1033954 1034739 786 pir:A70623 Mycobacterium tuberculosis 46.5 68.9 241 lpqU protein H37Rv Rv1022 lpqU 1085 4585 1034949 1036223 1275 sp:ENO_BACSU Bacillus subtilis eno 64.5 86.0 422 enolase (2-phosphoglycerate dehydratase)(2-phospho-D- glycerate hydro-lyase) 1086 4586 1036159 1036016 144 PIR:B72477 Aeropyrum pernix K1 APE2459 68.0 58.0 41 hypothetical protein 1087 4587 1036316 1036855 540 pir:C70623 Mycobacterium tuberculosis 31.9 55.0 191 hypothetical protein H37Rv Rv1024 1088 4588 1036900 1037445 546 pir:D70623 Mycobacterium tuberculosis 59.5 77.8 153 hypothetical protein H37Rv Rv1025 1089 4589 1037448 1038410 963 sp:GPPA_ECOLI Escherichia coli gppA 25.2 55.0 329 guanosine pentaphosphatase or exopolyphosphatase 1090 4590 1037481 1036498 984 1091 4591 1039650 1038721 930 sp:THD2_ECOLI Escherichia coli tdcB 30.3 64.7 314 threonine dehydratase 1092 4592 1039783 1039977 195 1093 4593 1039996 1040325 330 1094 4594 1040494 1040682 189 pir:B72287 Thermotoga maritima MSB8 46.3 74.1 56 hypothetical protein 1095 4595 1040925 1041917 993 sp:RHAR_ECOLI Escherichia coli rhaR 24.8 55.8 242 transcription activator of L-rhamnose operon 1096 4596 1042027 1042842 816 pir:F70893 Mycobacterium tuberculosis 57.8 80.1 282 hypothetical protein H37Rv Rv1072 1097 4597 1043236 1042850 387 1098 4598 1043747 1043298 450 gp:SCF55_39 Streptomyces coelicolor A3(2) 30.0 57.1 140 hypothetical protein SCF55.39 1099 4599 1044295 1043774 522 sp:GREA_ECOLI Escherichia coli greA 35.0 60.1 143 transcription elongation factor 1100 4600 1044959 1044477 483 pir:G70894 Mycobacterium tuberculosis 34.3 72.1 140 hypothetical protein H37Rv Rv1081c 1101 4601 1045158 1046030 873 pir:S44952 Streptomyces lincolnensis lmbE 31.7 56.3 300 lincomycin-production 1102 4602 1046073 1046390 318 1103 4603 1046610 1047707 1098 sp:AROG_CORGL Corynebacterium glutamicum 99.2 99.5 367 3-deoxy-D-arabino-heptulosonate-7- aroG phosphate synthase 1104 4604 1047452 1046820 633 1105 4605 1047827 1048501 675 sp:YARF_CORGL Corynebacterium glutamicum 96.0 97.3 97 hypothetical protein or undecaprenyl CCRC18310 pyrophosphate synthetase 1106 4606 1048356 1048529 174 SP:YARF_CORGL Corynebacterium glutamicum 100.0 100.0 28 hypothetical protein (Brevibacterium flavum) 1107 4607 1048525 1049043 519 1108 4608 1049385 1049068 318 1109 4609 1050362 1049427 936 sp:COAA_ECOLI Escherichia coli coaA 53.9 79.9 308 pantothenate kinase 1110 4610 1050624 1051925 1302 gsp:R97745 Brevibacterium flavum MJ-233 99.5 100.0 434 serine hydroxymethyl transferase glyA 1111 4611 1052021 1053880 1860 sp:PABS_STRGR Streptomyces griseus pabS 47.6 70.1 696 p-aminobenzoic acid synthase 1112 4612 1053880 1054602 723 1113 4613 1054859 1055722 864 1114 4614 1055032 1054640 393 1115 4615 1055783 1056319 537 gp:A01504_1 Alcaligenes faecalis ptcR 30.3 58.8 165 phosphinothricin resistance protin 1116 4616 1057200 1056322 879 sp:YBGK_ECOLI Escherichia coli ybgK 30.3 59.0 300 hypothetical protein 1117 4617 1057573 1058628 1056 1118 4618 1057868 1057200 669 sp:YBGJ_ECOLI Escherichia coli ybgJ 37.8 57.8 225 hypothetical protein 1119 4619 1058598 1057843 756 sp:LAMB_EMENI Emericella nidulans lamB 30.8 52.2 276 lactam utilization protein 1120 4620 1059214 1058624 591 sp:YCSH_BACSU Bacillus subtilis ycsH 40.6 81.2 165 hypothetical membrane protein 1121 4621 1059218 1059889 672 1122 4622 1059360 1059962 603 1123 4623 1060112 1060792 681 sp:YDHC_BACSU Bacillus subtilis ydhC 26.0 63.2 204 transcriptional regulator 1124 4624 1060869 1062146 1278 1125 4625 1063629 1062211 1419 sp:FUMH_RAT Rattus norvegicus (Rat) fumH 52.0 79.4 456 fumarate hydratase precursor 1126 4626 1063936 1064424 489 gp:AF048979_1 Rhodococcus erythropolis 32.7 65.4 159 NADH-dependent FMN IGTS8 dszD oxydoreductase 1127 4627 1064738 1064478 261 1128 4628 1065200 1064754 447 1129 4629 1065867 1065304 564 gp:SCAH10_16 Streptomyces coelicolor A3(2) 55.4 81.0 184 reductase StAH10.16 1130 4630 1066083 1067570 1488 sp:SOXA_RHOSO Rhodococcus sp. IGTS8 soxA 39.1 67.7 443 dibenzothiophene desulfurization enzyme A 1131 4631 1067570 1068649 1080 sp:SOXC_RHOSO Rhodococcus sp. IGTS8 soxC 25.8 51.3 372 dibenzothiophene desulfurization enzyme C (DBT sulfur dioxygenase) 1132 4632 1068649 1069845 1197 sp:SOXC_RHOSO Rhodococcus sp. IGTS8 soxC 28.9 61.6 391 dibenzothiophene desulfurization enzyme C (DBT sulfur dioxygenase) 1133 4633 1069692 1068913 780 1134 4634 1069808 1069119 690 1135 4635 1069959 1071134 1176 gp:ECO237695_3 Escherichia coli K12 ssuD 45.3 73.1 397 FMNH2-dependent aliphatic sulfonate monooxygenase 1136 4636 1072441 1071479 963 sp:GLPX_ECOLI Escherichia coli K12 glpX 44.3 75.7 325 glycerol metabolism 1137 4637 1072676 1073245 570 pir:B70897 Mycobacterium tuberculosis 27.5 56.4 211 hypothetical protein H37Rv Rv1100 1138 4638 1075241 1073340 1902 pir.H70062 Bacillus subtilis ywmD 31.3 66.1 227 hypothetical protein 1139 4639 1075357 1075641 285 1140 4640 1075553 1075329 225 gp:SCH24_37 Streptomyces coelicolor A3(2) 36.6 78.1 82 transmembrane efflux protein SCH24.37 1141 4641 1075909 1075667 243 sp:EX7S_ECOLI Escherichia coli K12 MG1655 40.3 67.7 62 exodeoxyribonuclease small subunit xseB 1142 4642 1077183 1075933 1251 sp:EX7L_ECOLI Escherichia coli K12 MG1655 30.0 55.6 466 exodeoxyribonuclease large subunit xseA 1143 4643 1077297 1078271 975 sp:LYTB_ECOLI Escherichia coli K12 lytB 50.2 78.8 311 penicillin tolerance 1144 4644 1077734 1077306 429 GSP:Y75421 Neisseria gonorrhoeae 33.0 47.0 131 polypeptides predicted to be useful antigens for vaccines and diagnostics 1145 4645 1079146 1078319 828 1146 4646 1080540 1079221 1320 sp:PERM_ECOLI Escherichia coli K12 perM 26.3 63.9 338 permease 1147 4647 1080965 1080786 180 1148 4648 1082708 1080972 1737 sp:NTPR_RAT Rattus norvegicus (Rat) SLC6A7 30.3 61.4 552 sodium-dependent proline ntpR transporter 1149 4649 1084183 1082951 1233 sp:CSP1_CORGL Corynebacterium glutamicum 29.9 60.0 412 major secreted protein PS1 protein (Brevibacterium flavum) ATCC precursor 17965 csp1 1150 4650 1084380 1085462 1083 sp:YYAF_BACSU Bacillus subtilis yyaF 70.1 88.6 361 GTP-binding protein 1151 4651 1085791 1086087 297 sp:VAPI_BACNO Dichelobacter nodosus intA 57.3 80.0 75 virulence-associated protein 1152 4652 1086096 1086917 822 sp:OTCA_PSEAE Pseudomonas aeruginosa argF 29.6 58.8 301 ornithine carbamoyltransferase 1153 4653 1087544 1087044 501 sp:YKKB_BACSU Bacillus subtilis 168 ykkB 39.2 69.9 143 hypothetical protein 1154 4654 1088293 1087664 630 gp:AF013288_1 Mus musculus RDH4 33.8 60.6 198 9-cis retinol dehydrogenase or oxidoreductase 1155 4655 1089740 1088535 1206 sp:YIS1_STRCO Streptomyces coelicolor 42.2 73.0 396 transposase/integrase (IS110) SC3C8.10 1156 4656 1090175 1093216 3042 sp:YEGE_ECOLI Escherichia coli K12 yegE 23.0 52.2 1153 hypothetical membrane protein 1157 4657 1093929 1094693 765 sp:NODC_RHIME Rhizobium meliloti nodC 22.8 47.1 259 N-acetylglucosaminyltransferase 1158 4658 1094693 1094911 219 1159 4659 1095052 1095384 333 1160 4660 1095677 1095387 291 pir:S43613 Corynebacterium glutamicum 82.5 93.8 97 transposase (insertion sequence ATCC 31831 IS31831) 1161 4661 1096093 1095719 375 pir:JC4742 Corynebacterium glutamicum 79.2 94.4 125 transposase (Brevibacterium lactofermentum) ATCC 13869 1162 4662 1096331 1096188 144 pir:JC4742 Corynebacterium glutamicum 87.5 95.8 48 transposase (Brevibacterium lactofermentum) ATCC 13869 1163 4663 1096471 1096331 141 1164 4664 1097111 1096746 366 1165 4665 1097229 1097726 498 1166 4666 1097750 1098592 843 sp:MORA_PSEPU Pseudomonas putida M10 norA 37.5 66.3 264 oxidoreductase or morpyine-6- dehydrogenase (naloxone reductase) 1167 4667 1098609 1098929 321 sp:DC4C_ACICA Acinetobacter calcoaceticus 33.3 63.9 108 4-carboxymuconolactone dc4c decarboxlyase 1168 4668 1099088 1099750 663 1169 4669 1099209 1099015 195 1170 4670 1099768 1099115 654 gp:AF058302_19 Streptomyces roseofulvus frnS 34.9 66.4 146 frenolicin gene cluster protein involved in frenolicin biosynthetic 1171 4671 1099917 1101653 1737 gp:SPU59234_3 Synechococcus sp. PCC 7942 48.1 78.5 563 biotin carboxylase accC 1172 4672 1102043 1102639 597 1173 4673 1102695 1103192 498 1174 4674 1103180 1103524 345 1175 4675 1103951 1104103 153 1176 4676 1104923 1105561 639 1177 4677 1106058 1104103 1956 sp:YT15_MYCTU Mycobacterium tuberculosis 57.9 80.3 655 hypothetical protein H37Rv Rv0959 1178 4678 1107381 1106086 1296 sp:BCHI_RHOSH Rhodobacter sphaeroides ATCC 27.7 52.6 329 magnesium chelatase subunit 17023 bchl 1179 4679 1107560 1108201 642 gp:AMU73808_1 Amycolatopsis methanolica pgm 33.8 62.5 160 2,3-PDG dependent phosphoglycerate mutase 1180 4680 1108201 1108905 705 pir:A70577 Mycobacterium tuberculosis 38.2 60.7 262 hypothetical protein H37Rv Rv2133c 1181 4681 1108993 1109754 762 gp:STMBCPA_1 Streptomyces hygroscopicus 29.4 59.3 248 carboxyphosphonoenolpyruvate SF1293 BcpA phosphonomutase 1182 4682 1109792 1111432 1641 sp:TLRC_STRFR Streptomyces fradiae tlrC 31.7 54.1 593 tyrosin resistance ATP-binding protein 1183 4683 1111820 1111425 396 sp:Y06C_MYCTU Mycobacterium tuberculosis 29.4 66.9 136 hypothetical protein H37Rv Rv2923c 1184 4684 1111889 1112230 342 sp:PHNA_ECOLI Escherichia coli K12 MG1655 55.0 82.0 111 alkylphosphonate uptake protein phnA 1185 4685 1112957 1112484 474 sp:YXAD_BACSU Bacillus subtilis 168 yxaD 32.1 62.7 134 transcriptional regulator 1186 4686 1113102 1114319 1218 gp:SPN7367_1 Streptococcus pneumoniae 22.6 59.4 367 multi-drug resistance efflux pump pmrA 1187 4687 1114486 1115793 1308 pir:S43613 Corynebacterium glutamicum 99.5 99.8 436 transposase (insertion sequence (Brevibacterium lactofermentum) IS31831) ATCC 31831 1188 4688 1116905 1115832 1074 gp:RFAJ3152_2 Ruminococcus flavefaciens 43.9 73.4 376 cysteine desulphurase cysteine desulphurase gene 1189 4689 1117744 1116908 837 sp:NADC_MYCTU Mycobacterium tuberculosis 42.1 68.9 283 nicotinate-nucleotide pyrophosphorylase 1190 4690 1118932 1117751 1182 pir:E69663 Bacillus subtilis nadA 49.3 77.6 361 quinolinate synthetase A 1191 4691 1119727 1119086 642 gp:SC5B8_7 Streptomyces coelicolor 37.0 60.9 235 DNA hydrolase SC5B8.07 1192 4692 1120205 1120804 600 gp:AE001961_5 Deinococcus radiodurans R1 23.4 54.7 192 hypothetical membrane protein DR1112 1193 4693 1121432 1120833 600 gp:SC3A7_8 Streptomyces coelicolor 36.0 66.4 214 hypothetical protein SC3A7.08 1194 4694 1121809 1121468 342 sp:YBDF_ECOLI Escherichia coli K12 MG1655 41.7 74.1 108 hypothetical protein ybdF 1195 4695 1122606 1121818 789 gp:AAA21740_1 Escherichia coli K12 lplA 30.1 60.7 216 lipoate-protein ligase A 1196 4696 1123051 1123461 411 sp:PHNB_ECOLI Escherichia coli K12 phnB 29.7 60.8 148 alkylphosphonate uptake protein and C-P lyase activity 1197 4697 1124826 1123534 1293 sp:PCAK_PSEPU Pseudomonas putida pcaK 28.8 64.3 420 transmembrane transport protein or 4-hydroxybenzoate transporter 1198 4698 1126020 1124836 1185 sp:PHHY_PSEAE Pseudomonas aeruginosa phhy 40.8 68.6 395 p-hydroxybenzoate hydroxylase (4- hydroxybenzoate 3- monooxygenase) 1199 4699 1126422 1127009 588 pir:A69859 Bacillus subtilis 168 ykoE 36.7 69.6 191 hypothetical membrane protein 1200 4700 1127013 1128350 1338 sp:YJJK_ECOLI Escherichia coli yjjK 24.8 47.6 532 ABC transporter ATP-binding protein 1201 4701 1128350 1129102 753 pir:G69858 Bacillus subtilis 168 ykoC 25.6 61.6 250 hypothetical membrane protein 1202 4702 1129102 1129632 531 1203 4703 1129655 1130704 1050 sp:CHAA_ECOLI Escherichia coli chaA 33.3 69.0 339 Ca2+/H+antiporter ChaA 1204 4704 1130721 1131428 708 pir:C75001 Pyrococcus abyssi Orsay 28.4 57.6 236 hypothetical protein PAB1341 1205 4705 1132123 1131401 723 sp:YWAF_BACSU Bacillus subtilis ywaF 27.6 61.1 221 hypothetical membrane protein 1206 4706 1134472 1132133 2340 sp:UVRA_THETH Thermus thermophilus unrA 35.5 58.7 946 excinuclease ABC subunit A 1207 4707 1134561 1135055 495 sp:TPX_MYCTU Mycobacterium tuberculosis 57.3 81.7 164 thioredoxin peroxidase H37Rv tpx 1208 4708 1135476 1135691 216 1209 4709 1136833 1135058 1776 1210 4710 1137891 1136938 954 sp:YEDI_ECOLI Escherichia coli yedL 39.9 72.0 318 hypothetical membrane protein 1211 4711 1137960 1138859 900 gp:SCF76_2 Streptomyces coelicolor A3(2) 34.0 49.0 282 oxidoreductase or thiamin biosynthesis protein 1212 4712 1138880 1139245 366 1213 4713 1139196 1139492 297 1214 4714 1139357 1139617 261 1215 4715 1140021 1139635 387 1216 4716 1140861 1140028 834 sp:CTR2_PENVA Penaeus vannamei 28.8 51.3 271 chymotrypsin Bll 1217 4717 1141245 1140901 345 sp:ARC2_ECOLI Escherichia coli 43.2 72.1 111 arsenate reductase (arsenical pump modifier) 1218 4718 1141273 1142472 1200 sp:YYAD_BACSU Bacillus subtilis yyaD 23.5 62.4 340 hypothetical membrane protein 1219 4719 1143015 1142479 537 pir:F70559 Mycobacterium tuberculosis 43.5 71.4 147 hypothetical protein H37Rv Rv1632c 1220 4720 1143739 1143026 714 pir:F70555 Mycobacterium tuberculosis 35.8 62.9 221 hypothetical protein H37Rv Rv1157c 1221 4721 1144118 1146028 1911 sp:TYPA_ECOLI Escherichia coli K12 typA 46.3 76.7 614 GTP-binding protein (tyrosine phsphorylated protein A) 1222 4722 1146097 1147602 1506 pir:F70874 Mycobacterium tuberculosis 27.9 54.9 506 hypothetical protein H37Rv Rv1166 1223 4723 1147592 1148461 870 pir:B70875 Mycobacterium tuberculosis 38.7 61.9 315 hypothetical protein H37Rv Rv1170 1224 4724 1148445 1148882 438 1225 4725 1148953 1149267 315 sp:FER_STRGR Streptomyces griseus fer 78.6 91.3 103 ferredoxin [4Fe-4S] 1226 4726 1149279 1150379 1101 sp:AAT_BACSP Bacillus sp. strain YM-2 aat 25.9 52.9 397 aspartate aminotransferase 1227 4727 1150408 1151028 621 1228 4728 1151186 1152370 1185 1229 4729 1153263 1152373 891 gp:CGAJ4934_1 Corynebacterium glutamicum 100.0 100.0 229 tetrahydrodipicolinate succinylase or ATCC 13032 dapD succinylation of piperidine-2,6- dicarboxylate 1230 4730 1156537 1155875 663 1231 4731 1156902 1157669 768 pir:S60064 Corynebacterium glutamicum 100.0 100.0 211 hypothetical protein ATCC 13032 orf2 1232 4732 1157694 1158524 831 gp:SCP8_4 Streptomyces coelicolor A3(2) 59.0 69.0 273 dihydropteroate synthase dhpS 1233 4733 1158524 1159252 729 gp:MLU15180_14 Mycobacterium leprae u1756l 45.7 73.1 245 hypothetical protein 1234 4734 1159267 1159572 306 pir:G70609 Mycobacterium tuberculosis 31.3 67.7 99 hypothetical protein H37Rv Rv1209 1235 4735 1159635 1159799 165 gsp:W32443 Mycobacterium tuberculosis 72.3 91.5 47 antigen TbAAMK, useful in vaccines for prevention or treatment of tuberculosis 1236 4736 1159865 1160728 864 sp:MYRA_MICGR Micromonospora griseorubida 39.2 67.8 286 mycinamicin-resistance gene myrA 1237 4737 1162231 1160738 1494 sp:SCRB_PEDPE Pediococcus pentosaceus scrB 23.5 51.0 524 sucrose-6-phosphate hydrolase 1238 4738 1163605 1162379 1227 sp:GLGA_ECOLI Escherichia coli K12 MG1655 24.7 51.3 433 ADPglucose-starch(bacterial glgA glycogen) glucosyltransferase 1239 4739 1163702 1164916 1215 sp:GLGC_STRCO Streptomyces coelicolor A3(2) 61.0 81.8 400 glucose-1-phosphate glgC adenylyltransferase 1240 4740 1165612 1164974 639 sp:MDMC_STRMY Streptomyces mycarofaciens 25.8 62.4 93 methyltransferase MdmC RNA polymerase sigma factor 1241 4741 1165746 1166384 639 sp:RPOE_ECOLI Escherichia coli rpoE 27.3 57.2 194 (sigma-24); heat shock and oxidative stress 1242 4742 1166576 1167067 492 1243 4743 1167110 1167577 468 pir:C70508 Mycobacterium tuberculosis 45.5 73.2 112 hypothetical protein H37Rv Rv1224 1244 4744 1168711 1167587 1125 sp:MRP_ECOLI Escherichia coli mrp 43.6 72.0 257 ATPase 1245 4745 1169325 1168747 579 pir:B70509 Mycobacterium tuberculosis 60.4 83.8 154 hypothetical protein H37Rv Rv1231c 1246 4746 1170610 1169321 1290 pir:C70509 Mycobacterium tuberculosis 49.8 77.0 434 hypothetical protein H37Rv Rv1232c 1247 4747 1170672 1171187 516 pir:A70952 Mycobacterium tuberculosis 57.9 87.1 140 hypothetical protein H37Rv Rv1234 1248 4748 1171206 1171871 666 1249 4749 1172462 1171869 594 1250 4750 1176271 1172501 3771 prf:2306367A Corynebacterium glutamicum 99.4 99.8 1257 2-oxoglutarate dehydrogenase AJ12036 odhA 1251 4751 1180048 1176308 3741 sp:MDR2_CRIGR Cricetulus griseus (Chinese 28.8 60.4 1288 ABC transporter or multidrug hamster) MDR2 resistance protein 2 (P-glycoprotein 2) 1252 4752 1180837 1180121 717 pir:H70953 Mycobacterium tuberculosis 31.7 72.1 240 hypothetical protein H37Rv Rv1249c 1253 4753 1181675 1180872 804 sp:AROE_ECOLI Escherichia coli aroE 25.5 61.2 255 shikimate dehydrogenase 1254 4754 1181993 1183603 1611 sp:PNBA_BACSU Bacillus subtilis pnbA 35.7 64.7 501 para-nitrobenzyl esterase 1255 4755 1183607 1184257 651 1256 4756 1184280 1185155 876 1257 4757 1185742 1185218 525 1258 4758 1185825 1187039 1215 sp:TCR1_ECOLI Escherichia coli transposon 27.1 61.4 409 tetracycline resistance protein Tn1721 tetA 1259 4759 1187043 1188389 1347 sp:TCMA_STRGA Streptomyces glaucescens tcmA 32.4 64.2 444 metabolite export pump of tetracenomycin C resistance 1260 4760 1189822 1190526 705 1261 4761 1190622 1188388 2235 pir:S57636 Catharanthus roseus metE 45.2 72.2 774 5- methyltetrahydropteroyltriglutamate- -homocysteine S-methyltransferase 1262 4762 1191087 1191542 456 1263 4763 1192410 1193807 1398 gsp:Y29930 Nocardia asteroides strain KGB1 55.2 79.5 444 thiophene biotransformation protein 1264 4764 1193867 1194190 324 1265 4765 1194165 1195109 945 1266 4766 1195916 1195125 792 1267 4767 1195974 1197620 1647 1268 4768 1197624 1197815 192 1269 4769 1199543 1197990 1554 sp:CYDC_ECOLI Escherichia coli K12 MG1655 28.7 63.5 526 ABC transporter cydC 1270 4770 1201075 1199543 1533 sp:CYDD_ECOLI Escherichia coli K12 MG1655 29.4 58.4 551 ABC transporter cydD 1271 4771 1202088 1201090 999 gp:AB035086_2 Corynebacterium glutamicum 92.0 93.0 333 cytochrome bd-type menaquinol (Brevibacterium lactofermentum) oxidase subunit II cydB 1272 4772 1203632 1202094 1539 gp:AB035086_1 Corynebacterium glutamicum 99.6 99.0 512 cytochrome bd-type menaquinol (Brevibacterium lactofermentum) oxidase subunit I cydA 1273 4773 1206180 1203916 2265 sp:YEJH_ECOLI Escherichia coli K12 MG1655 26.4 55.0 402 helicase yejH 1274 4774 1206316 1206657 342 1275 4775 1207223 1206831 393 sp:MUTT_PROVU Proteus vulgaris mutT 36.9 65.6 98 mutator mutT protein ((7,8-dihydro- 8-oxoguanine-triphosphatase)(8- oxo-dGTPase)(dGTP pyrophosphohydrolase) 1276 4776 1207374 1208138 765 1277 4777 1209615 1208212 1404 sp:PROY_SALTY Salmonella typhimurium proY 51.3 85.0 433 proline-specific permease 1278 4778 1209934 1212129 2196 sp:DEAD_KLEPN Klebsiella pneumoniae CG43 48.1 74.3 643 DEAD box ATP-dependent RNA DEAD box ATP-dependent RNA helicase helicase deaD 1279 4779 1213115 1212429 687 prf:2323363BT Mycobacterium leprae 24.7 47.4 247 bacterial regulatory protein, tetR B1308_C2_181 family 1280 4780 1213269 1214858 1590 sp:PCPB_FLAS3 Sphingomonas flava pcpB 24.5 47.7 595 pentachlorophenol 4- monooxygenase 1281 4781 1214871 1215938 1068 sp:CLCE_PSESB Pseudomonas sp.B13 clcE 40.4 72.0 354 maleylacetate reductase 1282 4782 1215952 1216836 885 sp:CATA_ACICA Acinetobacter calcoaceticus 30.6 59.4 278 catechol 1,2-dioxygenase catA 1283 4783 1217374 1216904 471 1284 4784 1217982 1217443 540 pir:A70672 Mycobacterium tuberculosis 31.9 58.4 185 hypothetical protein H37Rv Rv2972c 1285 4785 1219895 1222996 3102 sp:SNF2_YEAST Saccharomyces cerevisiae 24.9 55.4 878 transcriptional regulator SNF2 1286 4786 1222905 1221841 1065 1287 4787 1222986 1223843 858 gp:SCO007731_6 Streptomyces coelicolor A3(2) 29.6 56.2 203 hypothetical protein orfZ 1288 4788 1223887 1225059 1173 pir:E70755 Mycobacterium tuberculosis 39.2 67.3 395 phosphoesterase H37Rv Rv1277 1289 4789 1225066 1227693 2628 sp:Y084_MYCTU Mycobacterium tuberculosis 29.7 59.6 915 hypothetical protein H37Rv Rv1278 1290 4790 1227587 1227282 306 1291 4791 1227657 1227340 318 1292 4792 1227863 1228636 774 gp:AB029896_1 Petroleum-degrading bacterium 37.3 64.6 220 esterase or lipase HD-1 hde 1293 4793 1228718 1229095 378 1294 4794 1229150 1229935 786 1295 4795 1229716 1229180 537 sp:ATOE_ECOLI Streptomyces coelicolor 37.7 69.7 122 short-chain fatty acids transporter SC1C2.14c atoE 1296 4796 1229995 1230480 486 sp:PECS_ERWCH Erwinia chrysanthemi recS 24.7 56.6 166 regulatory protein 1297 4797 1230610 1230831 222 1298 4798 1231432 1230914 519 1299 4799 1231730 1232479 750 sp:FNR_ECOLI Escherichia coli K12 MG1655 fnr 25.0 57.9 228 fumarate (and nitrate) reduction regulatory protein 1300 4800 1232603 1232836 234 sp:MERP_SHEPU Shewanella putrefaciens merP 33.3 66.7 81 mercuric transort protein periplasmic component precursor 1301 4801 1233007 1234881 1875 sp:ATZN_ECOLI Escherichia coli K12 MG1655 38.0 70.6 605 zinc-transporting ATPase Zn(II)- atzN translocating P-type ATPase 1302 4802 1234983 1235612 630 sp:RELA_VIBSS Vibrio sp. S14 relA 32.9 58.4 137 GTP pyrophosphokinase (ATP:GTP 3′-pyrophosphotransferase) (ppGpp synthetase I) 1303 4803 1238125 1236545 1581 gsp:R80504 Streptomyces lividans tap 26.6 49.3 601 tripeptidyl aminopeptidase 1304 4804 1242156 1241554 603 1305 4805 1242275 1242156 120 1306 4806 1243621 1243728 108 GSP:P61449 Corynebacterium glutamicum 95.0 98.0 24 homoserine dehydrogenase 1307 4807 1245201 1243942 1260 1308 4808 1245532 1244843 690 1309 4809 1246496 1245720 777 sp:NARI_BACSU Bacillus subtilis narl 45.0 69.6 220 nitrate reductase gamma chain 1310 4810 1247239 1246508 732 sp:NARJ_BACSU Bacillus subtilis narJ 30.3 63.4 175 nitrate reductase delta chain 1311 4811 1248791 1247199 1593 sp:NARH_BACSU Bacillus subtilis narH 56.6 83.4 505 nitrate reductase beta chain 1312 4812 1249851 1250444 594 PIR:D72603 Aeropyrum pernix K1 APE1291 36.0 48.0 137 hypothetical protein 1313 4813 1251545 1251817 273 PIR:B72603 Aeropyrum pernix K1 APE1289 36.0 55.0 83 hypothetical protein 1314 4814 1252537 1248794 3744 sp:NARG_BACSU Bacillus subtilis narG 46.9 73.8 1271 nitrate reductase alpha chain 1315 4815 1253906 1252557 1350 sp:NARK_ECOLI Escherichia coli K12 narK 32.8 67.9 461 nitrate extrusion protein 1316 4816 1254146 1254634 489 sp:CNX1_ARATH Arabidopsis thaliana CV cnx1 32.5 65.0 157 molybdopterin biosynthesis cnx1 protein (molybdenum cofactor biosynthesis enzyme cnx1) 1317 4817 1256602 1254737 1866 sp:PRTS_SERMA Serratia marcescens strain IFO- 21.1 45.9 738 extracellular serine protease 3046 prtS precurosor 1318 4818 1257067 1257750 684 1319 4819 1257858 1256851 1008 sp:Y0D3_MYCTU Mycobacterium tuberculosis 30.8 62.6 334 hypothetical membrane protein H37Rv Rv1841c 1320 4820 1259265 1257865 1401 sp:Y0D2_MYCTU Mycobacterium tuberculosis 31.6 60.2 472 hypothetical membrane protein H37Rv Rv1842c 1321 4821 1259989 1259429 561 gp:PPU242952_2 Pseudomonas putida mobA 27.5 52.3 178 molybdopterin guanine dinucleotide synthase 1322 4822 1261201 1259993 1209 sp:MOEA_ECOLI Mycobacterium tuberculosis 32.8 58.2 366 molybdoptein biosynthesis protein H37Rv Rv0438c moeA 1323 4823 1262818 1261688 1131 sp:CNX2_ARATH Arabidopsis thaliana cnx2 51.4 73.7 354 molybdopterin biosynthsisi protein Moybdenume (mosybdenum cofastor biosythesis enzyme) 1324 4824 1264610 1262886 1725 sp:ALKK_PSEOL Pseudomonas oleovorans 36.7 65.7 572 edium-chain fatty acid-CoA ligase 1325 4825 1265142 1267427 2286 sp:RHO_MICLU Micrococcus luteus rho 50.7 73.8 753 Rho factor 1326 4826 1265665 1266267 603 1327 4827 1266306 1265611 696 1328 4828 1266449 1265427 1023 1329 4829 1267430 1268503 1074 sp:RF1_ECOLI Escherichia coli K12 RF-1 41.9 71.9 363 peptide chain release factor 1 1330 4830 1268507 1269343 837 sp:HEMK_ECOLI Escherichia coli K12 31.1 57.9 280 protoporphyrinogen oxidase 1331 4831 1269040 1268267 774 1332 4832 1269396 1270043 648 sp:YD01_MYCTU Mycobacterium tuberculosis 62.3 86.0 215 hypothetical protein H37Rv Rv1301 1333 4833 1270047 1271192 1146 sp:RFE_ECOLI Escherichia coli K12 rfe 31.1 58.4 322 undecaprenyl-phosphate alpha-N- acetylglucosaminyltransferase 1334 4834 1271213 1271698 486 1335 4835 1271871 1272119 249 GPU:AB046112_1 Corynebacterium glutamicum 98.0 99.0 80 hypothetical protein atpl 1336 4836 1272340 1273149 810 sp:ATP6_ECOLI Escherichia coli K12 atpB 24.1 56.7 245 ATP synthase chain a (protein 6) 1337 4837 1273286 1273525 240 sp:ATPL_STRLI Streptomyces lividans atpL 54.9 85.9 71 H+-transporting ATP synthase lipid- binding protein. ATP synthase C chane 1338 4838 1273559 1274122 564 sp:ATPF_STRLI Streptomyces lividans atpF 27.8 66.9 151 H+-transporting ATP synthase chain b 1339 4839 1274131 1274943 813 sp:ATPD_STRLI Streptomyces lividans atpD 34.3 67.2 274 H+-transporting ATP synthase delta chain 1340 4840 1274975 1276648 1674 sp:ATPA_STRLI Streptomyces lividans atpA 66.9 88.4 516 H+-transporting ATP synthase alpha chain 1341 4841 1276708 1277682 975 sp:ATPG_STRLI Streptomyces lividans atpG 46.3 76.6 320 H+-transporting ATP synthase gamma chain 1342 4842 1277688 1279136 1449 sp:ATPB_CORGL Corynebacterium glutamicum 99.8 100.0 483 H+-transporting ATP synthase beta AS019 atpB chain 1343 4843 1279151 1279522 372 sp:ATPE_STRLI Streptomyces lividans atpE 41.0 73.0 122 H+-transporting ATP synthase epsilon chain 1344 4844 1279770 1280240 471 sp:Y02W_MYCTU Mycobacterium tuberculosis 38.6 67.4 132 hypothetical protein H37Rv Rv1312 1345 4845 1280270 1280959 690 sp:Y036_MYCTU Mycobacterium tuberculosis 70.0 85.7 230 hypothetical protein H37Rv Rv1321 1346 4846 1280967 1281251 285 GP:SC26G5_35 Streptomyces coelicolor A3(2) 45.0 56.0 95 putative ATP/GTP-binding protein 1347 4847 1281714 1281262 453 sp:YQJC_BACSU Bacillus subtilis yqjC 35.8 68.7 134 hypothetical protein 1348 4848 1281794 1282105 312 sp:YC20_MYCTU Mycobacterium tuberculosis 54.5 79.2 101 hypothetical protein H37Rv Rv1898 1349 4849 1282194 1283114 921 sp:YD24_MYCTU Mycobacterium tuberculosis 37.9 71.4 301 thioredoxin H37Rv Rv1324 1350 4850 1283324 1284466 1143 gp:ECO237695_3 Escherichia coli K12 ssuD 50.3 74.3 366 FMNH2-dependent aliphatic sulfonate monooxygenase 1351 4851 1284517 1285284 768 sp:SSUC_ECOLI Escherichia coli K12 ssuC 40.8 75.8 240 alphatic sulfonates transport permease protein 1352 4852 1285302 1286030 729 sp:SSUB_ECOLI Escherichia coli K12 ssuB 50.4 72.8 228 alphatic sulfonates transport permease protein 1353 4853 1286043 1286999 957 sp:SSUA_ECOLI Escherichia coli K12 ssuA 35.1 62.1 311 sulfonate binding protein precursor 1354 4854 1289473 1287281 2193 sp:GLGB_ECOLI Mycobacterium tuberculosis 46.1 72.7 710 1,4-alpha-glucan branching enzyme H37Rv Rv1326c glgB (glycogen branching enzyme) 1355 4855 1291007 1289514 1494 sp:AMY3_DICTH Dictyoglomus thermophilum 22.9 50.5 467 alpha-amylase amyC 1356 4856 1291026 1291373 348 1357 4857 1291699 1292577 879 sp:FEPC_ECOLI Escherichia coli K12 fepC 31.8 87.6 211 ferric enterobactin transport ATP- binding protein or ABC transport ATP-binding protein 1358 4858 1293222 1294025 804 pir:C70860 Mycobacterium tuberculosis 39.6 68.5 260 hypothetical protein H37Rv Rv3040c 1359 4859 1294151 1295206 1056 pir:H70859 Mycobacterium tuberculosis 43.1 70.0 367 hypothetical protein H37Rv Rv3037c 1360 4860 1295047 1294436 612 1361 4861 1295435 1296220 786 sp:FIXA_RHIME Rhizobium meliloti fixA 31.2 64.8 244 electron transfer flavoprotein beta- subunit 1362 4862 1296253 1297203 951 sp:FIXB_RHIME Rhizobium meliloti fixB 33.1 61.8 335 electron transfer flavoprotein alpha subunit for various dehydrogenases 1363 4863 1296479 1297093 615 1364 4864 1297212 1298339 1128 sp:NIFS_AZOVI Azotobacter vinelandii nifS 35.2 67.7 375 nitrogenase cofactor sythesis protein 1365 4865 1298653 1298342 312 1366 4866 1300145 1299000 1146 sp:Y4ME_RHISN Rhizobium sp. NGR234 plasmid 29.5 55.7 397 hypothetical protein pNGR234a y4mE 1367 4867 1300369 1300145 225 sp.Y4MF_RHISN Rhizobium sp. NGR234 plasmid 47.5 76.3 59 transcriptional regulator pNGR234a Y4mF 1368 4868 1300552 1301055 504 sp:YHBS_ECOLI Escherichia coli K12 MG1655 34.8 55.3 181 acetyltransferase yhbS 1369 4869 1301929 1300988 942 1370 4870 1303123 1301975 1149 1371 4871 1303299 1303694 396 1372 4872 1303829 1304923 1095 pir:C70858 Mycobacterium tuberculosis 61.8 80.9 361 tRNA (5-methylaminomethyl-2- H37Rv Rv3024c thiouridylate)-methyltransferase 1373 4873 1304536 1303883 654 1374 4874 1304932 1305921 990 pir:B70857 Mycobacterium tuberculosis 33.7 66.0 332 hypothetical protein H37Rv Rv3015c 1375 4875 1307384 1305924 1461 sp:TCMA_STRGA Streptomyces glaucescens tcmA 30.2 65.8 500 tetracenomycin C resistance and export protin 1376 4876 1308196 1307462 735 1377 4877 1308330 1310369 2040 sp:DNLJ_RHOMR Rhodothermus marinus dnlJ 42.8 70.6 677 DNA ligase (polydeoxyribonucleotide synthase [NAD+] 1378 4878 1311097 1310435 663 pir:H70856 Mycobacterium tuberculosis 40.0 70.9 220 hypothetical protein H37Rv Rv3013 1379 4879 1311320 1311616 297 sp:GATC_STRCO Streptomyces coelicolor A3(2) 53.0 64.0 97 glutamyl-tRNA(Gln) gatC amidotransferase subunit C 1380 4880 1311625 1313115 1491 sp:GATA_MYCTU Mycobacterium tuberculosis 74.0 83.0 484 glutamyl-tRNA(Gln) H37Rv gatA amidotransferase subunit A 1381 4881 1313270 1314118 849 sp:VIUB_VIBVU Vibrio vulnificus viuB 28.1 54.0 263 vibriobactin utilization protein / iron- chelator utilization protein 1382 4882 1314775 1314470 306 gp:SCE6_24 Streptomyces coelicolor A3(2) 46.9 79.2 96 hypothetical membrane protein SCE6.24 1383 4883 1315013 1316083 1071 sp:PFP_AMYME Amycolatopsis methanolica pfp 54.8 77.9 358 pyrophosphate-fructose 6- phosphate 1-phosphotransrefase 1384 4884 1315954 1315325 630 1385 4885 1316338 1317444 1107 sp:CCPA_BACME Bacillus megaterium ccpA 31.4 31.4 328 glucose-resistance amylase regulator (catabolite control protein) 1386 4886 1317434 1319005 1572 sp:RBSA_ECOLI Escherichia coli K12 rbsA 44.7 76.2 499 ripose transport ATP-binding protein 1387 4887 1319005 1319976 972 sp:RBSC_ECOLI Escherichia coli K12 MG1655 45.6 76.9 329 high affinity ribose transport protein rbsC 1388 4888 1320001 1320942 942 sp:RBSB_ECOLI Escherichia coli K12 MG1655 45.9 77.7 305 periplasmic ribose-binding protein rbsB 1389 4889 1320952 1321320 369 sp:RBSD_ECOLI Escherichia coli K12 MG1655 41.7 68.4 139 high affinity ribose transport protein rbsD 1390 4890 1321476 1322111 636 sp:YIW2_YEAST Saccharomyces cerevisiae 31.0 58.0 200 hypothetical protein YIR042c 1391 4891 1322393 1323406 1014 gp:SCF34_13 Streptomyces coelicolor 31.4 60.2 354 iron-siderophore binding lipoprotein SCF34.13c 1392 4892 1323533 1324537 1005 sp:NTCI_RAT Rattus norvegicus (Rat) NTCI 35.8 61.9 268 Na-dependent bile acid transporter 1393 4893 1324778 1326256 1479 gsp:W61467 Staphylococcus aureus WHU 29 43.1 71.8 485 RNA-dependent amidotransferase B ratB 1394 4894 1326378 1327049 672 sp:F4RE_METJA Methanococcus jannaschii 32.6 61.1 172 putative F420-dependent NADH MJ1501 f4re reductase 1395 4895 1330967 1329891 1077 sp:YQJG_ECOLI Escherichia coli K12 yqjG 39.8 66.9 317 hypothetical protein 1396 4896 1331102 1331875 774 pir:A70672 Mycobacterium tuberculosis 39.3 62.4 234 hypothetical protein H37Rv Rv2972c 1397 4897 1331953 1333008 1056 pir:H70855 Mycobacterium tuberculosis 27.4 52.6 325 hypothetical membrane protein H37Rv Rv3005c 1398 4898 1333424 1333188 237 1399 4899 1335280 1333442 1839 gp:AJ012293_1 Corynebacterium glutamicum 99.2 99.4 613 dihydroxy-acid dehydratase ATCC 13032 ilvD 1400 4900 1335975 1335412 564 pir:G70855 Mycobacterium tuberculosis 33.3 68.6 105 hypothetical protein H37Rv Rv3004 1401 4901 1337567 1336095 1473 sp:YILV_CORGL Corynebacterium glutamicum 100.0 100.0 62 hypothetical membrane protein ATCC 13032 yilV 1402 4902 1338609 1338379 231 GP:SSU18930_263 Sulfolobus solfataricus 45.0 55.0 66 hypothetical protein 1403 4903 1342072 1342677 606 1404 4904 1342457 1341960 498 sp:NRTD_SYNP7 Synechococcus sp. nrtD 50.9 80.8 167 nitrate transport ATP-binding potein 1405 4905 1342727 1342461 267 sp:MALK_ENTAE Enterobacter aerogenes 46.0 78.2 87 maltose/maltodextrin transport ATP- (Aerobacter aerogenes) malK binding protein 1406 4906 1343675 1342794 882 sp:NRTA_ANASP Anabaena sp strain PCC 7120 28.1 56.8 324 nitrate transporter protein nrtA 1407 4907 1344018 1344464 447 1408 4908 1344440 1344808 369 1409 4909 1344935 1345420 486 sp:DIM6_STRCO Streptomyces coelicolor 39.4 73.2 142 actinorhodin polyketide dimerase 1410 4910 1345486 1346439 954 sp:CZCD_ALCEU Ralstonia eutropha czcD 39.1 72.7 304 cobalt-zinc-cadimium resistance protein 1411 4911 1345487 1345335 153 1412 4912 1346331 1345642 690 1413 4913 1346458 1348272 1815 sp:Y686_METJA Methanococcus jannaschii 22.9 53.7 642 hypothetical protein 1414 4914 1348334 1350076 1743 1415 4915 1350855 1352444 1590 gsp:Y22646 Brevibacterium flavum serA 99.8 100.0 530 D-3-phosphoglycerate dehydrogenase 1416 4916 1352053 1351727 327 SP:YEN1_SCHPO Schizosaccharomyces pombe 29.0 52.0 105 hypothetical serine-rich protein SPAC11G7.01 1417 4917 1352585 1353451 867 1418 4918 1355601 1354540 1062 1419 4919 1355689 1357554 1866 pir:T03476 Rhodobacter capsulatus strain 32.9 63.1 620 hypothetical protein SB1003 1420 4920 1356452 1356853 402 1421 4921 1357557 1358210 654 1422 4922 1358259 1359062 804 sp:HPCE_ECOLI Escherichia coli C hpcE 33.3 59.2 228 homoprotocatechiuate catabolism bifunctional isomerase/decarboxylase [includes: 2-hydroxyhepta-2,4-diene-1,7-dioate isomerase(hhdd isomerase); 5- carboxymethyl-2-oxo-hex-3-ene-1,7- dioate decarboxylase(opet decarboxylase)] 1423 4923 1359052 1359669 618 sp:UBIG_ECOLI Escherichia coli K12 23.4 55.7 192 methyltransferase or 3- demethylubiquinone-9 3-O- methyltransferase 1424 4924 1361295 1360168 1128 sp:DHBC_BACSU Bacillus subtilis dhbC 38.0 70.4 371 isochorismate synthase 1425 4925 1361361 1362848 1488 sp:SYE_BACSU Bacillus subtilis gltX 37.3 69.7 485 glutamyl-tRNA synthetase 1426 4926 1363138 1362926 213 gp:SCJ33_10 Streptomyces coelicolor A3(2) 77.0 90.0 67 transcriptional regulator 1427 4927 1363657 1363142 516 1428 4928 1364253 1363732 522 1429 4929 1364915 1365256 342 1430 4930 1364960 1364340 621 1431 4931 1365180 1364878 303 1432 4932 1365396 1365217 180 1433 4933 1365808 1366137 330 1434 4934 1367293 1367505 213 1435 4935 1368070 1367888 183 1436 4936 1368078 1368395 318 1437 4937 1368400 1369551 1152 1438 4938 1369551 1369874 324 1439 4939 1371637 1369877 1761 sp:THIC_BACSU Bacillus subtilis thiA or thiC 65.1 81.0 599 thiamin biosynthesis protein 1440 4940 1372326 1371979 348 1441 4941 1372601 1373131 531 1442 4942 1373798 1373929 132 GSP:Y37857 Chlamydia trachomatis 61.0 74.0 44 lipoprotein 1443 4943 1374556 1375491 936 1444 4944 1375776 1373350 2427 sp:PHS1_RAT Rattus norvegicus (Rat) 44.2 74.0 797 glycogen phosphorylase 1445 4945 1375987 1375805 183 1446 4946 1376088 1375933 156 1447 4947 1377555 1376149 1407 sp:YRKH_BACSU Bacillus subtilis yrkH 25.4 52.8 299 hypothetical protein 1448 4948 1378415 1377666 750 sp:Y441_METJA Methanococcus jannaschii Y441 25.4 64.8 256 hypothetical membrane protein 1449 4949 1378942 1378466 477 1450 4950 1379003 1379566 564 sp:SPOT_ECOLI Escherichia coli K12 spoT 29.8 60.1 178 guanosine 3′,5′-bis(diphosphate) 3′- pyrophosphatase 1451 4951 1380259 1379555 705 sp:ICLR_ECOLI Escherichia coli K12 iclR 26.1 60.7 257 acetate repressor protein 1452 4952 1380440 1381882 1443 sp:LEU2_ACTTI Actinoplanes teichomyceticus 68.1 87.5 473 3-isopropylmalate dehydratase large leu2 subunit 1453 4953 1381902 1382492 591 sp:LEUD_SALTY Salmonella typhimurium 67.7 89.2 195 3-isopropylmalate dehydratase small subunit 1454 4954 1382819 1382502 318 1455 4955 1383798 1382845 954 gp:MLCB637_35 Mycobacterium tuberculosis 45.9 71.4 294 mutator mutT protein ((7,8-dihydro- H37Rv MLCB637.35c 8-oxoguanine-triphosphatase)(8- oxo-dGTPase)(dGTP pyrophosphohydrolase) 1456 4956 1383930 1384085 156 1457 4957 1384130 1385125 996 sp:GPDA_BACSU Bacillus subtilis gpdA 45.0 72.2 331 NAD(P)H-dependent dihydroxyacetone phosphate reductase 1458 4958 1385153 1386232 1080 sp:DDLA_ECOLI Escherichia coli K12 MG1655 40.4 67.4 374 D-alanine-D-alanine ligase ddlA 1459 4959 1387270 1386293 978 1460 4960 1387332 1388324 993 sp:THIL_ECOLI Escherichia coli K12 thiL 32.2 57.6 335 thiamin-phosphate kinase 1461 4961 1388312 1389073 762 sp:UNG_MOUSE Mus musculus ung 38.8 59.6 245 uracil-DNA glycosylase precursor 1462 4962 1389208 1390788 1581 sp:Y369_MYCGE Mycoplasma genitalium (SGC3) 23.1 56.3 568 hypothetical protein MG369 1463 4963 1390796 1392916 2121 sp:RECG_ECOLI Escherichia coli K12 recG 35.4 60.0 693 ATP-dependent DNA helicase 1464 4964 1391961 1391638 324 GSP:Y75303 Neisseria meningitidis 31.0 48.0 108 polypeptides predicted to be useful antigens for vaccines and diagnostics 1465 4965 1392939 1393151 213 sp:BCCP_PROFR Propionibacterium freudenreichii 38.8 67.2 67 biotin carboxyl carrier protein subsp. Shermanii 1466 4966 1393154 1393735 582 sp:YHHF_ECOLI Escherichia coli K12 yhhF 37.1 63.5 167 methylase 1467 4967 1393742 1394221 480 sp:KDTB_ECOLI Escherichia coli K12 MG1655 42.6 78.7 155 lipopolysaccharide core biosynthesis kdtB protein 1468 4968 1394854 1395933 1080 1469 4969 1394894 1395097 204 GSP:Y75358 Neisseria gonorrhoeae 67.0 74.0 65 Neisserial polypeptides predicted to be useful antigens for vaccines and diagnostics 1470 4970 1395549 1394800 750 sp:GLNQ_BACST Bacillus stearothermophilus 56.4 78.6 252 ABC transporter or glutamine ABC glnQ transporter, ATP-binding protein 1471 4971 1396410 1395568 843 sp:NOCM_AGRT5 Agrobacterium tumefaciens 32.7 75.0 220 nopaline transport protein nocM 1472 4972 1397421 1396561 861 sp:GLNH_ECOLI Escherichia coli K12 MG1655 27.4 59.0 234 glutamine-binding protein precursor glnH 1473 4973 1397662 1398468 807 1474 4974 1399534 1398557 978 pir:H69160 Methanobacterium 28.6 60.3 322 hypothetical membrane protein thermoautotrophicum MTH465 1475 4975 1400926 1401333 408 1476 4976 1400940 1400185 756 sp:VINT_BPL54 Bacteriophage L54a vinT 26.9 52.5 223 phage integrase 1477 4977 1401333 1402076 744 1478 4978 1402272 1402703 432 1479 4979 1402874 1402368 507 1480 4980 1403128 1403991 864 1481 4981 1403997 1404215 219 1482 4982 1404885 1404694 192 pir:S60890 Corynebacterium glutamicum 88.5 96.2 26 insertion element (IS3 related) orf2 1483 4983 1406174 1405320 855 1484 4984 1407109 1406999 111 PIR:S60890 Corynebacterium glutamicum 89.0 97.0 37 hypothetical protein 1485 4985 1407535 1407167 369 1486 4986 1407873 1407559 315 1487 4987 1409023 1408703 321 1488 4988 1409802 1409428 375 1489 4989 1411011 1410064 948 1490 4990 1411424 1411119 306 1491 4991 1412000 1411437 564 1492 4992 1412351 1412572 222 1493 4993 1412916 1412626 291 1494 4994 1413745 1416459 2715 sp:DPO1_MYCTU Mycobacterium tuberculosis 56.3 80.8 896 DNA polymerase I polA 1495 4995 1417883 1416462 1422 sp:CMCT_NOCLA Streptomyces lactamdurans 33.8 67.8 456 cephamycin export protein cmcT 1496 4996 1417962 1418870 909 gp:SCJ9A_15 Streptomyces coelicolor A3(2) 41.3 65.4 283 DNA-binding protein SCJ9A.15c 1497 4997 1418876 1419748 873 sp:MORA_PSEPU Pseudomonas putida morA 46.5 76.1 284 morphine-6-dehydrogenase 1498 4998 1420036 1419878 159 1499 4999 1420724 1420071 654 sp:YAFE_ECOLI Streptomyces coelicolor 31.9 58.3 163 hypothetical protein SCH5.13 yafE 1500 5000 1421099 1422556 1458 sp:RS1_ECOLI Escherichia coli K12 rpsA 39.5 71.4 451 30S ribosomal protein S1 1501 5001 1422571 1421096 1476 1502 5002 1425279 1425878 600 sp:YACE_BRELA Brevibacterium lactofermentum 80.5 93.9 195 hypothetical protein ATCC 13869 yacE 1503 5003 1426257 1427354 1098 1504 5004 1427957 1427376 582 1505 5005 1428049 1427804 246 1506 5006 1428290 1429246 957 1507 5007 1429159 1428224 936 sp:IUNH_CRIFA Crithidia fasciculata iunH 61.9 81.0 310 inosine-uridine preferring nucleoside hypolase (purine nucleosidase) 1508 5008 1430642 1429194 1449 sp:QACA_STAAU Staphylococcus aureus 23.6 53.8 517 aniseptic resistance protein 1509 5009 1431579 1430659 921 sp:RBSK_ECOLI Escherichia coli K12 rbsK 35.5 67.6 293 ribose kinase 1510 5010 1432612 1431575 1038 sp:ASCG_ECOLI Escherichia coli K12 ascG 30.0 65.6 337 criptic asc operon repressor, ranscription regulator 1511 5011 1432750 1433547 798 1512 5012 1434105 1436201 2097 sp:UVRB_STRPN Streptococcus pneumoniae 57.4 83.3 671 excinuclease ABC subunit B plasmid pSB470 uvrB 1513 5013 1436335 1436775 441 sp:Y531_METJA Methanococcus jannaschii 33.6 59.2 152 hypothetical protein MJ0531 1514 5014 1437249 1436869 381 sp:YTFH_ECOLI Escherichia coli K12 ytfH 38.8 80.2 121 hypothetical protein 1515 5015 1437356 1438201 846 sp:YTFG_ECOLI Escherichia coli K12 ytfG 53.8 77.1 279 hypothetical protein 1516 5016 1439343 1440026 684 1517 5017 1440560 1438212 2349 pir:H70040 Bacillus subtilis yvgS 23.2 47.2 839 hypothetical protein 1518 5018 1441586 1440675 912 gp:SC9H11_26 Streptomyces coelicolor A3(2) 32.7 68.0 150 hypothetical protein SC9H11.26c 1519 5019 1442392 1441793 600 sp:YCBL_ECOLI Escherichia coli K12 ycbL 30.4 58.4 214 hydrolase 1520 5020 1442487 1445333 2847 sp:UVRA_ECOLI Escherichia coli K12 uvrA 56.2 80.6 952 excinuclease ABC subunit A 1521 5021 1444115 1443810 306 PIR:JQ0406 Micrococcus luteus 40.0 57.0 100 hypothetical protein 1246 (uvrA region) 1522 5022 1445393 1444944 450 PIR:JQ0406 Micrococcus luteus 31.0 47.0 142 hypothetical protein 1246 (uvrA region) 1523 5023 1446158 1446874 717 1524 5024 1447446 1445323 2124 1525 5025 1447792 1448358 567 sp:IF3_RHOSH Rhodobacter sphaeroides infC 52.5 78.2 179 translation initiation factor IF-3 1526 5026 1448390 1448581 192 sp:RL35_MYCFE Mycoplasma fermentans 41.7 76.7 60 50S ribosomal protein L35 1527 5027 1448645 1449025 381 sp:RL20_PSESY Pseudomonas syringae pv. 75.0 92.7 117 50S ribosomal protein L20 syringae 1528 5028 1449940 1449119 822 1529 5029 1450126 1450692 567 1530 5030 1450918 1451820 903 sp:UGPA_ECOLI Escherichia coli K12 MG1655 33.2 71.6 292 sn-glycerol-3-phosphate transport ugpA system permease protein 1531 5031 1451820 1452653 834 sp:UGPE_ECOLI Escherichia coli K12 MG1655 33.3 70.4 270 sn-glycerol-3-phosphate transport upgE system protein 1532 5032 1452758 1454071 1314 sp:UGPB_ECOLI Escherichia coli K12 MG1655 26.6 57.6 436 sn-glycerol-3-phosphate transport ugpB system permease proein 1533 5033 1454115 1455338 1224 sp:UGPC_ECOLI Escherichia coli K12 MG1655 44.0 71.3 393 sn-glycerol-3-phosphate transport ugpC ATP-binding protein 1534 5034 1454350 1454102 249 PIR:E72756 Aeropyrum pernix K1 APE0042 47.0 56.0 74 hypothetical protein 1535 5035 1456066 1455350 717 sp:GLPQ_BACSU Bacillus subtilis glpQ 26.2 50.0 244 glycerophosphoryl diester phosphodiesterase 1536 5036 1456355 1456948 594 sp:TRMH_ECOLI Escherichia coli K12 MG1655 34.0 71.2 153 tRNA(guanosine-2′-0-)- trmH methlytransferase 1537 5037 1457047 1458066 1020 sp:SYFA_BACSU Bacillus subtilis 168 syfA phenylalanyl-tRNA synthetase alpha chain 1538 5038 1458133 1460616 2484 sp:SYFB_ECOLI Escherichia coli K12 MG1655 42.6 71.7 343 phenylalanyl-tRNA synthetase beta syfB chain 1539 5039 1458966 1458196 771 1540 5040 1461157 1462128 972 sp:ESTA_STRSC Streptomyces scabies estA 26.5 55.1 363 esterase 1541 5041 1462134 1463516 1383 sp:MDMB_STRMY Streptomyces mycarofaciens 30.0 56.3 423 macrolide 3-O-acyltransferase mdmB 1542 5042 1463533 1463934 402 1543 5043 1464083 1465123 1041 gp:AF005242_1 Corynebacterium glutamicum 98.3 99.1 347 N-acetylglutamate-5-semialdehyde ASO19 argC dehydrogenase 1544 5044 1465210 1466373 1164 sp:ARGJ_CORGL Corynebacterium glutamicum 99.5 99.7 388 glutamate N-acetyltransferase ATCC 13032 argJ 1545 5045 1467376 1468548 1173 sp:ARGD_CORGL Corynebacterium glutamicum 99.0 99.2 391 acetylornithine aminotransferase ATCC 13032 argD 1546 5046 1470211 1471413 1203 sp:ASSY_CORGL Corynebacterium glutamicum 99.5 99.5 401 argininosuccinate synthetase ASO19 argG 1547 5047 1471362 1470154 1209 1548 5048 1471477 1472907 1431 gp:AF048764_1 Corynebacterium glutamicum 83.3 90.0 478 argininosuccinate lyase ASO19 argH 1549 5049 1472977 1474119 1143 1550 5050 1474119 1475693 1575 1551 5051 1475683 1476294 612 1552 5052 1476343 1476519 177 sp:YCAR_ECOLI Escherichia coli K12 ycaR 48.0 72.0 50 hypothetical protein 1553 5053 1476550 1477809 1260 sp:SYY1_BACSU Bacillus subtilis syy1 48.4 79.6 417 tyrosyl-tRNA synthase (tyrosine- tRNA ligase) 1554 5054 1478393 1477929 465 sp:Y531_METJA Methanococcus jannaschii 26.9 64.4 149 hypothetical protein MJ0531 1555 5055 1478892 1478503 390 1556 5056 1483475 1483335 141 PIR:F81737 Chlamydia muridarum Nigg 71.0 75.0 42 hypothetical protein TC0129 1557 5057 1483996 1483724 273 GSP:Y35814 Chlamydia pneumoniae 61.0 66.0 84 hypothetical protein 1558 5058 1484675 1486027 1353 sp:IF2_BORBU Borrelia burgdorferi IF2 36.3 67.0 182 translation initiation factor IF-2 1559 5059 1486042 1487025 984 sp:YZGD_BACSU Bacillus subtilis yzgD 29.6 60.1 311 hypothetical protein 1560 5060 1487032 1487193 162 1561 5061 1487238 1488056 819 sp:YQXC_BACSU Bacillus subtilis yqxC 38.5 69.6 260 hypothetical protein 1562 5062 1488146 1489018 873 sp:YFJB_HAEIN Mycobacterium tuberculosis 31.6 31.6 225 hypothetical protein H37Rv Rv1695 1563 5063 1489103 1490881 1779 sp:RECN_ECOLI Escherichia coli K12 recN 31.4 63.4 574 DNA repair protein 1564 5064 1490944 1492134 1191 pir:H70502 Mycobacterium tuberculosis 41.9 73.1 394 hypothetical protein H37Rv Rv1697 1565 5065 1492147 1493109 963 pir:A70503 Mycobacterium tuberculosis 30.4 68.1 313 hypothetical protein H37Rv Rv1698 1566 5066 1493513 1495174 1662 sp:PYRG_ECOLI Escherichia coli K12 pyrG 55.0 76.7 549 CTP synthase (UTP-ammonia ligase) 1567 5067 1495205 1495861 657 sp:YQKG_BACSU Bacillus subtilis yqkG 36.3 71.3 157 hypothetical protein 1568 5068 1495861 1496772 912 gp:AF093548_1 Staphylococcus aureus xerD 39.7 71.7 300 tyrosine recombinase 1569 5069 1498324 1496795 1530 sp:TLRC_STRFR Streptomyces fradiae tlrC 30.5 59.7 551 tyrosin resistance ATP-binding protein 1570 5070 1498863 1499645 783 gp:CCU87804_4 Caulobacter crescentus parA 44.6 73.6 258 chromosome partitioning protein or ATPase involved in active partitioning of diverse bacterial plasmids 1571 5071 1499931 1500695 765 sp:YPUG_BACSU Bacillus subtilis ypuG 28.3 64.5 251 hypothetical protein 1572 5072 1501471 1500911 561 1573 5073 1501710 1502576 867 gp:AF109156_1 Datisca glomerata tst 35.6 67.0 270 thiosulfate sulfurtransferase 1574 5074 1502634 1503176 543 sp:YPUH_BACSU Bacillus subtilis ypuH 33.1 65.7 172 hypothetical protein 1575 5075 1503483 1504238 756 sp:RLUB_BACSU Bacillus subtilis rluB 45.9 72.5 229 ribosomal large subunit pseudouridine synthase B 1576 5076 1504256 1504945 690 sp:KCY_BACSU Bacillus subtilis cmk 38.6 73.6 220 cytidylate kinase 1577 5077 1505017 1506573 1557 sp:YPHC_BACSU Bacillus subtilis yphC 42.8 74.0 435 GTP binding protein 1578 5078 1507327 1506662 666 1579 5079 1507902 1507405 498 1580 5080 1508729 1507917 813 sp:YX42_MYCTU Mycobacterium tuberculosis 36.2 67.2 232 methyltransferase Rv3342 1581 5081 1508813 1510366 1554 prf:2513302B Corynebacterium striatum M82B 29.7 60.1 499 ABC transporter tetA 1582 5082 1510366 1512132 1767 prf:2513302A Corynebacterium striatum M82B 31.2 56.3 602 ABC transporter tetB 1583 5083 1511667 1510843 825 1584 5084 151289 1512977 789 sp:YGIE_ECOLI Escherichia coli K12 ygiE 39.7 73.2 257 hypothetical membrane protein 1585 5085 1514505 1514693 189 1586 5086 1514527 1512980 1548 gp:AB029555_1 Bacillus subtilis ATCC 9372 25.7 61.5 499 Na+/H+ antiporter nhaG 1587 5087 1515159 1514974 186 1588 5088 1515396 1515815 420 1589 5089 1515782 1515408 375 sp:YCHJ_ECOLI Escherichia coli K12 o249#9 36.9 57.7 130 hypothetical protein ychJ 1590 5090 1516962 1515799 1164 pir:C69334 Archaeoglobus fulgidus AF0675 25.2 63.8 210 2-hydroxy-6-oxohepta-2,4-dienoate hydrolase 1591 5091 1517170 1519458 2289 sp:SECA_BACSU Bacillus subtilis secA 35.2 61.7 805 preprotein translocase SecA subunit 1592 5092 1519601 1520029 429 gp:AF173844_2 Mycobacterium smegmatis garA 75.8 93.2 132 signal transduction protein 1593 5093 1520190 1520945 756 sp:Y0DF_MYCTU Mycobacterium tuberculosis 41.9 74.4 234 hypothetical protein H37Rv Rv1828 1594 5094 1520957 1521589 633 sp:Y0DE_MYCTU Mycobacterium tuberculosis 30.8 63.2 133 hypothetical protein H37Rv Rv1828 1595 5095 1521771 1522343 573 sp:Y0DE_MYCTU Mycobacterium tuberculosis 71.4 84.3 178 hypothetical protein H37Rv Rv1828 1596 5096 1522941 1522432 510 1597 5097 1524500 1523052 1449 1598 5098 1525374 1525973 600 1599 5099 1525497 1524568 930 1600 5100 1526534 1525473 1062 sp:YHDP_BACSU Bacillus subtilis yhdP 33.9 69.0 342 hemolysin 1601 5101 1527913 1526534 1380 sp:YHDT_BACSU Bacillus subtilis yhdT 31.4 65.5 65 hemolysin 1602 5102 1527968 1528186 219 1603 5103 1529330 1527987 1344 gp:TTHERAGEN_1 Thermus thermophilus herA 41.2 69.5 374 DEAD box RNA helicase 1604 5104 1529486 1530220 735 sp:YD48_MYCTU Mycobacterium tuberculosis 34.3 66.1 245 ABC transporter ATP-binding protein H37Rv Rv1348 1605 5105 1531816 1530341 1476 gsp:W27613 Brevibacterium flavum 99.0 99.2 492 6-phosphogluconate dehydrogenase 1606 5106 1531933 1532394 462 pir:G70664 Mycobacterium tuberculosis 39.7 67.8 121 thioesterase H37Rv Rv1847 1607 5107 1532322 1532996 675 1608 5108 1533041 1533781 741 sp:NODI_RHIS3 Rhizobium sp. N33 nodl 39.6 68.1 235 nodulation ATP-binding proiten I 1609 5109 1533781 1534521 741 pir:E70501 Mycobacterium tuberculosis 43.1 76.3 232 hypothetical membrane protein H37Rv Rv1686c 1610 5110 1535401 1534529 873 sp:YFHH_ECOLI Escherichia coli K12 yfhH 26.7 63.9 277 transcriptional regulator 1611 5111 1536227 1535382 846 sp:PHNE_ECOLI Escherichia coli K12 phnE 29.9 63.4 281 phosphonates transport system permease protein 1612 5112 1537030 1536227 804 sp:PHNE_ECOLI Escherichia coli K12 phnE 27.2 62.3 268 phosphonates transport system permease protein 1613 5113 1537833 1537030 804 sp:PHNC_ECOLI Escherichia coli K12 phnC 44.8 72.0 250 phosphonates transport ATP-binding protein 1614 5114 1538759 1538968 210 1615 5115 1538919 1537870 1050 1616 5116 1539664 1538963 702 1617 5117 1541403 1539820 1584 sp:THID_SALTY Salmonella typhimurium thiD 47.3 70.2 262 phosphomethylpyrimidine kinase 1618 5118 1542922 1542119 804 sp:THIM_SALTY Salmonella typhimurium LT2 46.6 77.5 249 hydoxyethylthiazole kinase thiM 1619 5119 1544976 1546289 1314 pir:H70830 Mycobacterium tuberculosis 28.6 55.0 451 cyclopropane-fatty-acyl-phospholipid H37Rv ufaA1 synthase 1620 5120 1547692 1546307 1386 prf:2223339B Burkholderia cepacia Pc701 32.5 66.9 468 sugar transporter or 4-methyl-o- mopB phthalate/phthalate permease 1621 5121 1548440 1547967 474 prf:2120352B Thermus flavus AT-62 gpt 36.5 59.0 156 purine phosphoribosyltransferase 1622 5122 1548651 1549349 699 sp:YEBN_ECOLI Escherichia coli K12 yebN 39.8 68.5 206 hypothetical protein 1623 5123 1549403 1550398 996 gp:AF178758_2 Sinorhizobium sp. As4 arsB 23.3 54.6 361 arsenic oxyanion-translocation pump membrane subunit 1624 5124 1550469 1550951 483 1625 5125 1551545 1552237 693 gp:SC17_33 Streptomyces coelicolor A3(2) 62.2 83.8 222 hypothetical protein SC17.33 1626 5126 1552518 1553972 1455 gp:PSTRTETC1_6 Pseudomonas sp. R9 ORFA 51.8 83.6 469 sulfate permease 1627 5127 1553722 1553297 426 GP:PSTRTETC1_7 Pseudomonas sp. R9 ORFG 39.0 50.0 97 hypothetical protein 1628 5128 1554684 1554070 615 1629 5129 1554861 1555067 207 1630 5130 1555079 1554891 189 1631 5131 1555835 1555086 750 1632 5132 1556376 1556771 396 pir:A70945 Mycobacterium tuberculosis 71.8 87.3 110 hypothetical protein H37Rv Rv2050 1633 5133 1557823 1557014 810 prf:2317468A Schizosaccharomyces pombe 39.2 71.0 217 dolichol phosphate mannose dpm1 synthase 1634 5134 1559493 1557859 1635 sp:LNT_ECOLI Escherichia coli K12 Int 25.1 55.6 527 apolipoprotein N-acyltransferase 1635 5135 1560237 1559497 741 1636 5136 1561660 1560437 1224 gp:AF188894_1 Candida albicans lip 1 23.7 55.6 392 secretory lipase 1637 5137 1561780 1562553 774 pir:C70764 Mycobacterium tuberculosis 31.3 56.7 291 precorrin 2 methyltransferase H37Rv cobG 1638 5138 1563802 1562525 1278 sp:COBL_PSEDE Pseudomonas denitrificans 32.4 60.8 411 precorrin-6Y C5, 15- SC510 cobL methyltransferase 1639 5139 1563872 1564237 366 1640 5140 1564237 1564482 246 1641 5141 1565302 1564565 738 sp:YY12_MYCTU Mycobacterium tuberculosis 54.1 75.4 244 oxidoreductase H37Rv RV3412 1642 5142 1566438 1565302 1137 gp:AF014460_1 Streptococcus mutans LT11 36.1 61.3 382 dipeptidase or X-Pro dipeptidase pepQ 1643 5143 1566468 1567106 639 1644 5144 1569903 1567117 2787 sp:MTR4_YEAST Saccharomyces cerevisiae 26.5 55.7 1030 ATP-dependent RNA helicase YJL050W dob1 1645 5145 1570933 1569932 1002 sp:TATC_ECOLI Escherichia coli K12 tatC 28.7 62.7 268 sec-independent protein translocase protein 1646 5146 1571382 1571068 315 sp:YY34_MYCLE Mycobacterium leprae 44.7 69.4 85 hypothetical protein MLCB2533.27 1647 5147 1572486 1571506 981 sp:YY35_MYCTU Mycobacterium tuberculosis 31.9 61.2 317 hypothetical protein H37Rv Rv2095c 1648 5148 1573463 1572492 972 sp:YY36_MYCLE Mycobacterium leprae 32.4 64.8 324 hypothetical protein MLCB2533.25 1649 5149 1574915 1573491 1425 sp:YY37_MYCTU Mycobacterium tuberculosis 53.1 77.3 467 hypothetical protein H37Rv Rv2097c 1650 5150 1574957 1575205 249 1651 5151 1575136 1574945 192 pir:B70512 Mycobacterium tuberculosis 54.1 80.3 61 hypothetical protein H37Rv Rv2111c 1652 5152 1576947 1575406 1542 pir:C70512 Mycobacterium tuberculosis 48.6 74.2 516 hypothetical protein H37Rv Rv2112c 1653 5153 1577327 1577806 480 PIR:H72504 Aeropyrum pernix K1 APE2014 42.0 50.0 159 hypothetical protein 1654 5154 1578531 1576951 1581 prf:2422382Q Rhodococcus erythropolis arc 51.6 78.5 545 AAA family ATPase (chaperone-like function) 1655 5155 1579400 1578567 834 pir:S72844 Mycobacterium leprae pimT 57.3 79.0 281 protein-beta-aspartate methyltransferase 1656 5156 1580771 1579449 1323 gp:AF005050_1 Homo sapiens 38.1 67.2 436 aspartyl aminopeptidase 1657 5157 1580807 1581640 834 pir:B70513 Mycobacterium tuberculosis 45.4 71.4 269 hypothetical protein H37Rv Rv2119 1658 5158 1581851 1582114 264 sp:VAPI_BACNO Dichelobacter nodosus A198 40.6 72.5 69 virulence-associated protein vapl 1659 5159 1583481 1582273 1209 prf:2513299A Staphylococcus aureus norA23 21.8 61.0 385 quinolon resistance protein 1660 5160 1585490 1583913 1578 sp:ASPA_CORGL Corynebacterium glutamicum 99.8 99.8 526 aspartate ammonia-lyase (Brevibacterium flavum) MJ233 aspA 1661 5161 1586445 1585603 843 gp:AF050166_1 Corynebacterium glutamicum 96.8 97.5 281 ATP phosphoribosyltransferase ASO19 hisG 1662 5162 1587504 1586812 693 pir:H72277 Thermotoga maritima MSB8 30.8 63.1 195 beta-phosphoglucomutase TM1254 1663 5163 1591235 1587573 3663 sp:METH_ECOLI Escherichia coli K12 metH 31.6 62.4 1254 5-methyltetrahydrofolate- homocysteine methyltransferase 1664 5164 1591343 1591912 570 1665 5165 1592966 1591941 1026 sp:AHPF_XANCH Xanthomonas campestris ahpF 22.4 49.5 366 alkyl hydroperoxide reductase subunit F 1666 5166 1593337 1594512 1176 sp:ACR3_YEAST Saccharomyces cerevisiae 33.0 63.9 388 arsenical-resistance protein S288C YPR201W acr3 1667 5167 1594532 1594951 420 sp:ARSC_STAAU Staphylococcus aureus plasmid 32.6 64.3 129 arsenate reductase p1258 arsC 1668 5168 1595030 1595668 639 pir:G70964 Mycobacterium tuberculosis 47.2 75.6 123 arsenate reductase H37Rv arsC 1669 5169 1596221 1595844 378 1670 5170 1597460 1596249 1212 sp:SYC_ECOLI Escherichia coli K12 cysS 35.9 64.3 387 cysteinyl-tRNA synthetase 1671 5171 1598623 1597745 879 sp:BACA_ECOLI Escherichia coli K12 bacA 37.3 69.4 255 bacitracin resistance protein 1672 5172 1598667 1599614 948 prf.2214302F Agrobacterium tumefaciens 33.4 62.6 326 oxidoreductase mocA 1673 5173 1599679 1600677 999 pir:F70577 Mycobacterium tuberculosis 27.0 53.5 359 lipoprotein H37Rv lppL 1674 5174 1600692 1601804 1113 sp:PYRD_AGRAE Agrocybe aegerita ura 1 44.0 67.1 334 dihydroorotate dehydrogenase 1675 5175 1602281 1601931 351 1676 5176 1602660 1603466 807 1677 5177 1603520 1604629 1110 gp:PSESTBCBAD_1 Pseudomonas syringae tnpA 34.7 55.3 360 transposase 1678 5178 1605315 1604830 486 1679 5179 1605811 1605281 531 sp:YBHB_ECOLI Escherichia coli K12 ybhB 44.1 75.0 152 bio operon ORF I (biotin biosynthetic enzyme) 1680 5180 1605961 1606689 729 GSP:Y74829 Neisseria meningitidis 26.0 33.0 198 Neisserial polypeptides predicted to be useful antigens for vaccines and diagnostics 1681 5181 1607646 1608248 603 1682 5182 1607657 1605861 1797 prf:2513302A Corynebacterium striatum M82B 43.6 68.7 597 ABC transporter tetB 1683 5183 1609087 1609335 249 1684 5184 1609247 1607661 1587 prf:2513302B Corynebacterium striatum M82B 36.8 67.1 535 ABC transporter tetA 1685 5185 1610192 1609842 351 1686 5186 1610236 1610844 609 pir:JU0052 Streptomyces anulatus pac 32.4 56.4 56 puromycin N-acetyltransferase 1687 5187 1612238 1611150 1089 sp:ARGK_ECOLI Escherichia coli K12 argK 43.1 72.3 339 LAO(lysine, arginine, and ornithine)/AO (arginine and ornithine)transport system kinase 1688 5188 1614444 1612234 2211 sp:MUTB_STRCM Streptomyces cinnamonensis 72.2 87.5 741 methylmalonyl-CoA mutase alpha A3823.5 mutB subunit 1689 5189 1616298 1614451 1848 sp:MUTA_STRCM Streptomyces cinnamonensis 41.6 68.2 610 methylmalonyl-CoA mutase beta A3823.5 mutA subunit 1690 5190 1616578 1617300 723 sp:YS13_MYCTU Mycobacterium tuberculosis 39.7 70.1 224 hypothetical membrane protein H37Rv Rv1491c 1691 5191 1617398 1617994 597 1692 5192 1619616 1618321 1296 sp:YS09_MYCTU Mycobacterium tuberculosis 64.1 87.0 370 hypothetical membrane protein H37Rv Rv1488 1693 5193 1620106 1619672 435 pir:B70711 Mycobacterium tuberculosis 44.7 78.7 141 hypothetical membrane protein H37Rv Rv1487 1694 5194 1621009 1620167 843 gp:SCC77_24 Streptomyces coelicolor A3(2) 51.0 72.8 261 hypothetical protein SCC77.24 1695 5195 1621056 1621838 783 1696 5196 1622950 1621841 1110 sp:HEMZ_PROFR Propionibacterium freudenreichii 36.8 65.7 364 ferrochelatase subsp. Shermanii hemH 1697 5197 1624826 1623027 1800 sp:P54_ENTFC Streptococcus faecium 25.5 56.5 611 invasin 1698 5198 1625925 1625428 498 1699 5199 1626279 1629107 2829 pir:F70873 Mycobacterium tuberculosis 69.9 85.9 959 aconitate hydratase H37Rv acn 1700 5200 1629298 1629861 564 pir:E70873 Mycobacterium tuberculosis 54.6 81.6 174 transcriptional regulator H37Rv Rv1474c 1701 5201 1629913 1630668 756 pir:F64496 Methanococcus jannaschii 21.3 51.9 235 GMP synthetase MJ1575 guaA 1702 5202 1631329 1630667 663 gp:SCD82_4 Streptomyces coelicolor A3(2) 32.6 62.0 221 hypothetical protein SCD82.04c 1703 5203 1631660 1631926 267 pir:E64494 Methanococcus jannaschii 37.2 80.2 86 hypothetical protein MJ1558 1704 5204 1631745 1631353 393 1705 5205 1631933 1633324 1392 gp:AE002515_9 Neisseria meningitidis MC58 61.2 86.1 446 hypothetical protein NMB1652 1706 5206 1632588 1632109 480 GSP:Y38838 Neisseria gonorrhoeae ORF24 54.0 60.0 113 antigenic protein 1707 5207 1633137 1632682 456 GSP:Y38838 Neisseria gonorrhoeae 59.0 69.0 152 antigenic protein 1708 5208 1633566 1636241 2676 sp:ATA1_SYNY3 Synechocystis sp. PCC6803 42.6 73.2 883 cation-transporting ATPase P sll1614 pma1 1709 5209 1634563 1633781 783 1710 5210 1636732 1636244 489 gp:SC3D11_2 Streptomyces coelicolor A3(2) 35.8 58.3 120 hypothetical protein SC3D11.02c 1711 5211 1637081 1638442 1362 1712 5212 1639132 1638776 357 1713 5213 1639365 1639520 156 1714 5214 1639656 1639817 162 1715 5215 1639781 1640155 375 prf:2408488H Streptococcus thermophilus 43.0 73.8 107 host cell surface-exposed lipoprotein phage TP-J34 1716 5216 1640546 1641001 456 prf:2510491A Corynephage 304L int 34.4 60.4 154 integrase 1717 5217 1642674 1641046 1629 sp:YJJK_ECOLI Escherichia coli K12 yjjK 32.8 64.4 497 ABC transporter ATP-binding protein 1718 5218 1644218 1642743 1476 1719 5219 1645499 1644318 1182 sp:NANH_MICVI Micromonospora viridifaciens 51.9 72.4 387 sialidase ATCC 31146 nedA 1720 5220 1645661 1646368 708 gp:AF121000_8 Corynebacterium glutamicum 99 6 100.0 236 transposase (IS1628) 22243 R-plasmid pAG1 tnpB 1721 5221 1645821 1646063 243 GPU:AF164956_23 Corynebacterium glutamicum 64.0 72.0 37 transposase protein fragment TnpNC 1722 5222 1645861 1645601 261 GP:NT1TNIS_5 Plasmid NTP16 32.0 43.0 88 hypothetical protein 1723 5223 1646549 1647133 585 1724 5224 1647634 1647212 423 pir:B75015 Pyrococcus abyssi Orsay 32.7 70.1 107 dTDP-4-keto-L-rhamnose reductase PAB1087 1725 5225 1648097 1647651 447 pir:S72754 Mycobacterium leprae 63.8 85.2 149 nitrogen fixation protein MLCL536.24c nifU7 1726 5226 1648548 1648709 162 PIR:C72506 Aeropyrum pernix K1 APE2025 48.0 57.0 52 hypothetical protein 1727 5227 1649362 1648100 1263 pir.S72761 Mycobacterium leprae nifS 64.7 84.4 411 nitrogen fixation protein 1728 5228 1650122 1649367 756 gp:SCC22_4 Streptomyces coelicolor A3(2) 70.2 89.3 252 ABC transporter ATP-binding protein SCC22.04c 1729 5229 1651424 1650249 1176 pir:A70872 Mycobacterium tuberculosis 55.2 83.0 377 hypothetical protein H37Rv Rv1462 1730 5230 1652875 1651433 1443 sp:Y074_SYNY3 Synechocystis sp. PCC6803 41.0 73.0 493 ABC transporter slr0074 1731 5231 1653586 1652894 693 gp:SCC22_8 Streptomyces coelicolor A3(2) 46.1 71.4 217 DNA-binding protein SCC22.08c 1732 5232 1654043 1655671 1629 pir:F70871 Mycobacterium tuberculosis 36.3 67.8 518 hypothetical membrane protein H37Rv Rv1459c 1733 5233 1655681 1656700 1020 pir:S72783 Mycobacterium leprae 50.2 77.3 317 ABC transporter MLCL536.31 abc2 1734 5234 1656712 1657515 804 pir:S72778 Mycobacterium leprae 41.0 74.8 266 hypothetical protein MLCL536.32 1735 5235 1657677 1658675 999 pir:C70871 Mycobacterium tuberculosis 43.0 74.6 291 hypothetical protein H37Rv Rv1456c 1736 5236 1659496 1659140 357 1737 5237 1659508 1661136 1629 pir:C71156 Pyrococcus horikoshii PH0450 23.4 51.0 418 helicase 1738 5238 1661578 1662552 975 sp:QOR_ECOLI Escherichia coli K12 qor 37.5 70.9 323 quinone oxidoreductase 1739 5239 1663598 1662630 969 gp:NWCOXABC_3 Nitrobacter winogradskyi coxC 37.6 66.8 295 cytochrome o ubiquinol oxidase assembly factor / heme O synthase 1740 5240 1664403 1666502 2100 gp:AB023377_1 Corynebacterium glutamicum 100.0 100.0 675 transketolase ATCC 31833 tkt 1741 5241 1666673 1667752 1080 sp:TAL_MYCLE Mycobacterium leprae 62.0 85.2 358 transaldolase MLCL536.39 tal 1742 5242 1667764 1666601 1164 1743 5243 1667950 1669401 1452 gsp:W27612 Brevibacterium flavum 99.8 100.0 484 glucose-6-phosphate dehydrogenase 1744 5244 1669419 1670375 957 pir:A70917 Mycobacterium tuberculosis 40.6 71.7 318 oxppcycle protein (glucose 6- H37Rv Rv1446c opcA phosphate dehydrogenase assembly protein) 1745 5245 1670395 1671099 705 sp:SOL3_YEAST Saccharomyces cerevisiae 28.7 58.1 258 6-phosphogluconolactonase S288C YHR163W sol3 1746 5246 1671677 1671273 405 sp:SAOX_BACSN Bacillus sp. NS-129 35.2 57.8 128 sarcosine oxidase 1747 5247 1671723 1673123 1401 gp:AF126281_1 Rhodococcus erythropolis 24.6 46.6 500 transposase (IS1676) 1748 5248 1674105 1673266 840 gp:CGL007732_5 Corynebacterium glutamicum 100.0 100.0 205 sarcosine oxidase ATCC 13032 soxA 1749 5249 1677211 1677384 174 1750 5250 1678756 1678070 687 1751 5251 1679148 1680128 981 1752 5252 1681108 1680332 777 sp:TPIS_CORGL Corynebacterium glutamicum 99.2 99.6 259 triose-phosphate isomerase AS019 ATCC 13059 tpiA 1753 5253 1681263 1681670 408 SP.YCQ3_YEAST Saccharomyces cerevisiae 37.0 51.0 128 probable membrane protein YCR013c 1754 5254 1682404 1681190 1215 sp:PGK_CORGL Corynebacterium glutamicum 98.0 98.5 405 phosphoglycerate kinase AS019 ATCC 13059 pgk 1755 5255 1683625 1682624 1002 sp:G3P_CORGL Corynebacterium glutamicum 99.1 99.7 333 glyceraldehyde-3-phosphate AS019 ATCC 13059 gap dehydrogenase 1756 5256 1685097 1684117 981 pir:D70903 Mycobacterium tuberculosis 63.9 87.4 324 hypothetical protein H37Rv Rv1423 1757 5257 1686132 1685110 1023 sp:YR4O_MYCTU Mycobacterium tuberculosis 56.3 82.5 309 hypothetical protein H37Rv Rv1422 1758 5258 1687078 1686152 927 sp:YR39_MYCTU Mycobacterium tuberculosis 52.0 76.2 281 hypothetical protein H37Rv Rv1421 1759 5259 1689190 1687103 2088 sp:UVRC_PSEFL Synechocystis sp. PCC6803 34.4 61.5 701 excinuclease ABC subunit C uvrC 1760 5260 1689779 1689201 579 sp:YR35_MYCTU Mycobacterium tuberculosis 32.7 68.7 150 hypothetical protein H37Rv Rv1417 1761 5261 1690345 1689869 477 sp:RISB_ECOLI Escherichia coli K12 43.5 72.1 154 6,7-dimethyl-8-ribityllumazine synthase 1762 5262 1690694 1690921 228 GSP:Y83273 Bacillus subtilis 59.0 68.0 72 polypeptide encoded by rib operon 1763 5263 1690708 1691421 714 GSP:Y83272 Bacillus subtilis 26.0 48.0 217 riboflavin biosynthetic protein 1764 5264 1691012 1691347 336 GSP:Y83273 Bacillus subtilis 44.0 52.0 106 polypeptide encoded by rib operon 1765 5265 1691625 1690360 1266 gp:AF001929_1 Mycobacterium tuberculosis ribA 65.6 84.7 404 GTP cyclohydrolase II and 3,4- dihydroxy-2-butanone 4-phosphate synthase (riboflavin synthesis) 1766 5266 1692271 1691639 633 sp:RISA_ACTPL Actinobacillus 47.4 79.2 211 riboflavin synthase alpha chain pleuropneumoniae ISU-178 ribE 1767 5267 1693258 1692275 984 sp:RIBD_ECOLI Escherichia coli K12 ribD 37.3 62.7 365 riboflavin-specific deaminase 1768 5268 1693918 1693262 657 sp:RPE_YEAST Saccharomyces cerevisiae 43.6 73.1 234 ribulose-phosphate 3-epimerase S288C YJL121C rpe1 1769 5269 1695298 1693967 1332 sp:SUN_ECOLI Escherichia coli K12 sun 30.8 60.7 448 nucleolar protein NOL1/NOP2 (eukaryotes) family 1770 5270 1696443 1695499 945 sp:FMT_PSEAE Pseudomonas aeruginosa fmt 41.6 67.9 308 methionyl-tRNA formyltransferase 1771 5271 1696972 1696466 507 sp:DEF_BACSU Bacillus subtilis 168 def 44.7 72.7 150 polypeptide deformylase 1772 5272 1699147 1697084 2064 sp:PRIA_ECOLI Escherichia coli priA 22.9 46.3 725 primosomal protein n 1773 5273 1700397 1699177 1221 gsp:R80060 Brevibacterium flavum MJ-233 99.3 99.5 407 S-adenosylmethionine synthetase 1774 5274 1701767 1700508 1260 sp:DFP_MYCTU Mycobacterium tuberculosis 58.0 80.9 409 DNA/pantothenate metabolism H37Rv RV1391 dfp flavoprotein 1775 5275 1702322 1702032 291 sp:YD90_MYCTU Mycobacterium tuberculosis 70.4 87.7 81 hypothetical protein H37Rv Rv1390 1776 5276 1703037 1702411 627 pir:KIBYGU Saccharomyces cerevisiae guk1 39.8 74.7 186 guanylate kinase 1777 5277 1703308 1702991 318 pir:B70899 Mycobacterium tuberculosis 80.6 90.3 103 integration host factor H37Rv Rv1388 mIHF 1778 5278 1704350 1703517 834 sp:DCOP_MYCTU Mycobacterium tuberculosis 51.8 73.6 276 orotidine-5′-phosphate H37Rv uraA decarboxylase 1779 5279 1707697 1704359 3339 pir:SYECCP Escherichia coli carB 53.1 77.5 1122 carbamoyl-phosphate synthase large chain 1780 5280 1708884 1707706 1179 sp:CARA_PSEAE Pseudomonas aeruginosa 45.4 70.1 381 carbamoyl-phosphate synthase ATCC 15692 carA small chain 1781 5281 1710357 1709017 1341 sp:PYRC_BACCL Bacillus caldolyticus DSM 405 42.8 67.7 402 dihydroorotase pyrC 1782 5282 1711348 1710413 936 sp:PYRB_PSEAE Pseudomonas aeruginosa 48.6 79.7 311 aspartate carbamoyltransferase ATCC 15692 1783 5283 1711927 1711352 576 sp:PYRR_BACCL Bacillus caldolyticus DSM 405 54.0 80.1 176 phosphoribosyl transferase or pyrR pyrimidine operon regulatory protein 1784 5284 1712596 1713759 1164 sp:Y00R_MYCTU Mycobacterium tuberculosis 39.7 73.4 297 cell division inhibitor H37Rv Rv2216 1785 5285 1713830 1714306 477 1786 5286 1714299 1714760 462 1787 5287 1714741 1714950 210 1788 5288 1716062 1715382 681 sp:NUSB_BACSU Bacillus subtilis nusB 33.6 69.3 137 N utilization substance protein B (regulation of rRNA biosynthesis by transcriptional antitermination) 1789 5289 1716692 1716132 561 sp:EFP_BRELA Brevibacterium lactofermentum 97.9 98.4 187 elongation factor P ATCC 13869 efp 1790 5290 1717868 1716780 1089 gp:AF124600_4 Corynebacterium glutamicum 99.5 100.0 217 cytoplasmic peptidase AS019 pepQ 1791 5291 1719032 1717938 1095 gp:AF124600_3 Corynebacterium glutamicum 98.6 99.7 361 3-dehydroquinate synthase AS019 aroB 1792 5292 1719598 1719107 492 gp:AF124600_2 Corynebacterium glutamicum 100.0 100.0 166 shikimate kinase AS019 aroK 1793 5293 1721381 1720971 411 sp:LEP3_AERHY Aeromonas hydrophila tapD 35.2 54.9 142 type IV prepilin-like protein specific leader peptidase 1794 5294 1721725 1721423 303 gp:SC1A2_22 Streptomyces coelicolor A3(2) 45.8 68.7 83 bacterial regulatory protein, arsR SC1A2.22 family 1795 5295 1721780 1722853 1074 gp:AF109162_2 Corynebacterium diphtheriae 35.9 73.2 340 ABC transporter hmuU 1796 5296 1722807 1722202 606 1797 5297 1722870 1723826 957 pir:A75169 Pyrococcus abyssi Orsay 23.6 50.7 373 iron(III) ABC transporter, PAB0349 periplasmic-binding protein 1798 5298 1723826 1724578 753 sp:FHUC_BACSU Bacillus subtilis 168 fhuC 38.3 71.7 230 ferrichrome transport ATP-binding protein 1799 5299 1725439 1724612 828 pir:D70660 Mycobacterium tuberculosis 50.0 60.0 259 shikimate 5-dehydrogenase H37Rv aroE 1800 5300 1726625 1725459 1167 pir:E70660 Mycobacterium tuberculosis 41.8 70.1 395 hypothetical protein H37Rv Rv2553c 1801 5301 1727170 1726625 546 pir:F70660 Mycobacterium tuberculosis 52.8 69.6 161 hypothetical protein H37Rv Rv2554c 1802 5302 1730048 1727385 2664 sp:SYA_THIFE Thiobacillus ferrooxidans ATCC 43.3 71.8 894 alanyl-tRNA synthetase 33020 alaS 1803 5303 1731542 1730166 1377 sp:Y0A9_MYCTU Mycobacterium tuberculosis 65.4 84.8 454 hypothetical protein H37Rv Rv2559c 1804 5304 1732822 1731599 1224 1805 5305 1734811 1732988 1824 sp:SYD_MYCLE Mycobacterium leprae aspS 71.1 89.2 591 aspartyl-tRNA synthetase 1806 5306 1735056 1735946 891 sp:Y0BQ_MYCTU Mycobacterium tuberculosis 46.1 74.1 297 hypothetical protein H37Rv Rv2575 1807 5307 1738679 1736004 2676 sp:AMYH_YEAST Saccharomyces cerevisiae 26.1 53.6 839 glucan 1,4-alpha-glucosidase S288C YIR019C sta1 1808 5308 1740569 1738713 1857 sp:YHGE_BACSU Bacillus subtilis yhgE 23.1 54.0 742 phage infection protein 1809 5309 1741219 1740572 648 1810 5310 1741313 1741906 594 gp:SCE68_13 Streptomyces coelicolor A3(2) 29.2 62.0 192 transcriptional regulator SCE68.13 1811 5311 1741893 1742606 714 1812 5312 1742701 1743813 1113 gp:SCE15_13 Streptomyces coelicolor A3(2) 72.8 88.1 371 oxidoreductase SCE15.13c 1813 5313 1743843 1743968 126 1814 5314 1744025 1744519 495 sp:SLFA_PSEAE Pseudomonas aeruginosa PAO1 37.1 77.6 116 NADH-dependent FMN reductase slfA 1815 5315 1744884 1746230 1347 sp:SDHL_ECOLI Escherichia coli K12 sdaA 46.8 71.4 462 L-serine dehydratase 1816 5316 1746728 1747588 861 1817 5317 1747918 1746233 1686 prf:2423362A Enterococcus casseliflavus glpO 28.4 53.9 598 alpha-glycerolphosphate oxidase 1818 5318 1749276 1747990 1287 sp:SYH_STAAU Staphylococcus aureus 43.2 72.2 421 histidyl-tRNA synthetase SR17238 hisS 1819 5319 1749963 1749325 639 gp:CJ11168X3_127 Campylobacter jejuni 40.3 62.1 211 hydrolase NCTC11168 Cj0809c 1820 5320 1750427 1750933 507 prf:2313309A Streptomyces chrysomallus 35.4 61.1 175 cyclophilin sccypB 1821 5321 1750964 1751200 237 1822 5322 1751497 1752051 555 gp:AF038651_4 Corynebacterium glutamicum 98.4 100.0 128 hypothetical protein ATCC 13032 orf4 1823 5323 1752186 1752527 342 1824 5324 1754894 1752615 2280 gp:AF038651_3 Corynebacterium glutamicum 99.9 99.9 760 GTP pyrophosphokinase ATCC 13032 rel 1825 5325 1755479 1754925 555 gp:AF038651_2 Corynebacterium glutamicum 99.5 100.0 185 adenine phosphoribosyltransferase ATCC 13032 apt 1826 5326 1755748 1755599 150 gp:AF038651_1 Corynebacterium glutamicum 98.0 98.8 49 dipeptide transport system ATCC 13032 dciAE 1827 5327 1757228 1755486 1743 sp:Y0BG_MYCTU Mycobacterium tuberculosis 30.7 60.9 558 hypothetical protein H37Rv Rv2585c 1828 5328 1758797 1757589 1209 sp:SECF_ECOLI Escherichia coli K12 secF 25.9 57.2 332 protein-export membrane protein 1829 5329 1759707 1760336 630 1830 5330 1760734 1758803 1932 prf:2313285A Rhodobacter capsulatus secD 24.4 52.0 616 protein-export membrane protein 1831 5331 1761367 1761005 363 sp:Y0BD_MYCLE Mycobacterium leprae 39.6 66.0 106 hypothetical protein MLCB1259.04 1832 5332 1762498 1761419 1080 sp:RUVB_ECOLI Escherichia coli K12 ruvB 55.3 81.9 331 holliday junction DNA helicase 1833 5333 1763134 1762517 618 sp:RUVA_MYCLE Mycobacterium leprae ruvA 45.2 74.3 210 holliday junction DNA helicase 1834 5334 1763839 1763177 663 sp:RUVC_ECOLI Escherichia coli K12 ruvC 35.6 63.3 180 crossover junction endodeoxyribonuclease 1835 5335 1764742 1763990 753 sp.YEBC_ECOLI Escherichia coli K12 ORF246 49.2 78.4 250 hypothetical protein yebC 1836 5336 1765860 1765015 846 sp:TESB_ECOLI Escherichia coli K12 tesB 38.5 68.6 283 acyl-CoA thiolesterase 1837 5337 1765969 1766442 474 gp:SC10A5_9 Streptomyces coelicolor A3(2) 31.5 61.3 111 hypothetical protein SC10A5.09c 1838 5338 1766948 1766487 462 pir:H70570 Mycobacterium tuberculosis 38.2 61.2 170 hypothetical protein H37Rv Rv2609c 1839 5339 1768030 1766948 1083 sp:GPI3_YEAST Saccharomyces cerevisiae 21.7 49.3 414 hexosyltransferase or N- S288C spt14 acetylglucosaminyl- phosphatidylinositol biosynthetic protein 1840 5340 1768996 1768034 963 gp:SCL2_16 Streptomyces coelicolor A3(2) 46.4 67.8 295 acyltransferase SCL2.16c 1841 5341 1769678 1769022 657 pir:C70571 Mycobacterium tuberculosis 48.2 78.0 78 CDP-diacylglycerol-glycerol-3- H37Rv Rv2612c pgsA phosphate phosphatidyltransferase 1842 5342 1770340 1769681 660 pir:D70571 Mycobacterium tuberculosis 54.6 78.4 194 histidine triad (HIT) family protein H37Rv Rv2613c 1843 5343 1772384 1770327 2058 sp:SYT2_BACSU Bacillus subtilis thrZ 42.0 68.9 647 threonyl-tRNA synthetase 1844 5344 1773863 1772658 1206 sp:YWBN_BACSU Bacillus subtilis ywbN 34.3 61.8 400 hypothetical protein 1845 5345 1773881 1774444 564 1846 5346 1774438 1773893 546 1847 5347 1775191 1774457 735 1848 5348 1777269 1777646 378 1849 5349 1777444 1778037 594 1850 5350 1779508 1778102 1407 1851 5351 1780168 1779554 615 1852 5352 1780905 1780507 399 1853 5353 1781585 1781019 567 sp:PUAC_STRLP Streptomyces anulatus pac 36.3 64.2 190 puromycin N-acetyltransferase 1854 5354 1781705 1782790 1086 1855 5355 1783281 1784381 1101 1856 5356 1784080 1783382 699 1857 5357 1785473 1782894 2580 1858 5358 1786844 1785732 1113 1859 5359 1788829 1786907 1923 1860 5360 1789080 1789562 483 1861 5361 1789580 1789768 189 1862 5362 1789746 1790057 312 1863 5363 1790889 1790461 429 1864 5364 1791842 1792438 597 sp:AFUC_ACTPL Actinobacillus 28.7 28.7 202 ferric transport ATP-binding protein pleuropneumoniae afuC 1865 5365 1792428 1793426 999 1866 5366 1793654 1793496 159 1867 5367 1793714 1794820 1107 1868 5368 1795202 1795621 420 1869 5369 1795591 1796181 591 gp:AF088896_20 Zymomonas mobilis dfp 27.1 66.7 129 pantothenate metabolism flavoprotein 1870 5370 1796186 1797049 864 1871 5371 1797350 1797769 420 1872 5372 1797969 1797850 120 1873 5373 1798757 1798023 735 1874 5374 1799182 1799406 225 1875 5375 1799473 1800366 894 1876 5376 1800604 1800449 156 1877 5377 1800834 1801307 474 1878 5378 1801344 1802096 753 1879 5379 1802577 1802155 423 1880 5380 1802733 1803419 687 1881 5381 1803465 1803893 429 1882 5382 1804134 1804598 465 1883 5383 1804629 1804865 237 1884 5384 1804919 1805599 681 1885 5385 1805727 1806686 960 1886 5386 1806917 1807396 480 1887 5387 1807433 1808113 681 1888 5388 1808137 1808421 285 1889 5389 1808458 1808832 375 1890 5390 1809761 1810372 612 sp:TNP2_ECOLI Escherichia coli tnpR 51.1 78.0 186 transposon TN21 resolvase 1891 5391 1810541 1811545 1005 1892 5392 1811564 1811938 375 1893 5393 1812215 1812691 477 sp:PVH1_YEAST Saccharomyces cerevisiae 29.3 51.8 164 protein-tyrosine phosphatase S288C YIR026C yvh1 1894 5394 1812881 1813606 726 1895 5395 1812882 1812460 423 1896 5396 1813780 1814517 738 gp:SCA32WHIH_6 Streptomyces coelicolor A3(2) 34.3 65.7 216 sporulation transcription factor whiH 1897 5397 1814863 1815651 789 1898 5398 1815673 1816128 456 1899 5399 1816451 1816636 186 1900 5400 1817132 1817803 672 1901 5401 1817803 1818219 417 1902 5402 1818460 1818774 315 1903 5403 1818798 1819166 369 1904 5404 1819954 1819748 207 1905 5405 1822382 1820181 2202 pir:C72285 Thermotoga maritima MSB8 22.6 55.2 545 hypothetical protein TM1189 1906 5406 1822577 1824322 1746 1907 5407 1824371 1824589 219 1908 5408 1824784 1824927 144 1909 5409 1825606 1825178 429 1910 5410 1826024 1826557 534 PIR:S60891 Corynebacterium glutamicum 63.0 75.0 166 hypothetical protein 1911 5411 1826644 1825751 894 pir:S60890 Corynebacterium glutamicum 87.9 95.6 298 insertion element (IS3 related) orf2 1912 5412 1826937 1826644 294 pir:S60889 Corynebacterium glutamicum 72.3 84.2 101 insertion element (IS3 related) orf1 1913 5413 1829900 1829688 213 1914 5414 1830765 1832063 1299 1915 5415 1832167 1834044 1878 sp:RECJ_ERWCH Erwinia chrysanthemi recJ 24.0 50.6 622 single-stranded-DNA-specific exonuclease 1916 5416 1834928 1834149 780 1917 5417 1836675 1838324 1650 pir:T13302 Streptococcus phage phi-O1205 31.8 64.3 381 primase ORF13 1918 5418 1838349 1842137 3789 1919 5419 1842235 1842681 447 1920 5420 1842804 1843337 534 1921 5421 1843518 1845356 1839 sp:Y018_MYCPN Mycoplasma pneumoniae ATCC 22.1 44.7 620 helicase 29342 yb95 1922 5422 1845483 1845857 375 1923 5423 1845872 1846207 336 pir:T13144 Bacteriophage N15 gene57 36.7 64.2 109 phage N15 protein gp57 1924 5424 1846698 1846333 366 1925 5425 1847315 1847932 618 1926 5426 1847938 1848474 537 1927 5427 1848509 1849036 528 1928 5428 1848988 1849785 798 1929 5429 1849781 1849966 186 1930 5430 1850035 1850406 372 1931 5431 1850415 1849978 438 1932 5432 1851049 1850474 576 1933 5433 1851220 1852440 1221 gp:SPAP J760_2 Schizosaccharomyces pombe 28.7 49.8 422 actin binding protein with SH3 SPAP J76O.02c domains 1934 5434 1851473 1852324 852 1935 5435 1852479 1853873 1395 1936 5436 1854261 1854854 594 1937 5437 1855058 1855237 180 1938 5438 1855532 1856788 1257 gp:SC5C7_14 Streptomyces coelicolor 23.6 52.5 347 ATP/GTP binding protein SC5C7.14 1939 5439 1856885 1858738 1854 1940 5440 1858763 1860727 1965 sp:CLPA_ECOLI Escherichia coli K12 clpA 30.2 61.0 630 ATP-dependent Clp proteinase ATP- binding subunit 1941 5441 1860752 1861225 474 1942 5442 1861320 1861475 156 1943 5443 1861842 1861519 324 1944 5444 1862088 1862399 312 1945 5445 1862945 1865299 2355 sp:PCRA_STAAU Staphylococcus aureus SA20 21.4 45.9 693 ATP-dependent helicase pcrA 1946 5446 1865265 1865822 558 1947 5447 1865842 1866219 378 1948 5448 1866328 1866792 465 1949 5449 1866832 1867095 264 1950 5450 1867098 1867874 777 gp:SCH17_7 Streptomyces coelicolor A3(2) 25.9 47.8 224 hypothetical protein SCH17.07c 1951 5451 1867886 1868587 702 prf:2514444Y Bacteriophage phi-C31 gp52 31.7 61.5 208 deoxynucleotide monophosphate kinase 1952 5452 1868895 1868671 225 1953 5453 1871092 1868927 2166 1954 5454 1871373 1871101 273 1955 5455 1877886 1871380 6507 1956 5456 1878312 1879400 1089 prf:2403350A Corynebacterium glutamicum 99.2 99.7 363 type II 5-cytosoine ATCC 13032 cgllM methyltransferase 1957 5457 1879412 1880485 1074 pir:A55225 Corynebacterium glutamicum 99.7 99.7 358 type II restriction endonuclease ATCC 13032 cgllR 1958 5458 1883990 1882470 1521 1959 5459 1884936 1884220 717 1960 5460 1885230 1887047 1818 gp:SC1A2_16 Streptomyces coelicolor A3(2) 24.6 45.8 504 hypothetical protein SC1A2.16c 1961 5461 1887405 1887590 186 1962 5462 1888038 1887688 351 gp:AE001973_4 Deinococcus radiodurans 46.7 70.0 90 SNF2/Rad54 helicase-related DR1258 protein 1963 5463 1889094 1888231 864 pir:T13226 Lactobacillus phage phi-gle 33.1 56.4 163 hypothetical protein Rorf232 1964 5464 1889530 1889859 330 1965 5465 1891707 1890028 1680 gp:AF188935_16 Bacillus anthracis pX02-16 20.7 47.9 537 hypothetical protein 1966 5466 1893037 1891832 1206 1967 5467 1894680 1893388 1293 1968 5468 1897231 1894739 2493 1969 5469 1899158 1897374 1785 sp:CLPB_ECOLI Escherichia coli clpB 25.3 52.5 724 endopeptidase Clp ATP-binding chain B 1970 5470 1899853 1899233 621 1971 5471 1900916 1899804 1113 1972 5472 1901911 1901066 846 1973 5473 1901975 1902955 981 1974 5474 1902883 1902005 879 1975 5475 1903028 1903225 198 1976 5476 1905878 1903113 2766 pir:S23647 Homo sapiens numA 20.1 49.1 1004 nuclear mitotic apparatus protein 1977 5477 1906572 1905973 600 1978 5478 1907914 1906664 1251 1979 5479 1908660 1907965 696 1980 5480 1909498 1908785 714 1981 5481 1910508 1909501 1008 1982 5482 1912300 1910642 1659 1983 5483 1913820 1912333 1488 1984 5484 1914371 1913973 399 1985 5485 1916233 1914725 1509 1986 5486 1916374 1916733 360 1987 5487 1916944 1917165 222 1988 5488 1917640 1917329 312 1989 5489 1918208 1917564 645 1990 5490 1919461 1918703 759 1991 5491 1920194 1919646 549 1992 5492 1921276 1920347 930 1993 5493 1925390 1925695 306 1994 5494 1925682 1926038 357 1995 5495 1926010 1921547 4464 pir:T03099 Sus scrofa domestica 23.2 49.2 1408 submaxillary apomucin 1996 5496 1926837 1926259 579 1997 5497 1928189 1927245 945 1998 5498 1928211 1928381 171 sp:MTE1_ECOLI Escherichia coli ecoR1 42.6 65.6 61 modification methylase 1999 5499 1928534 1928908 375 2000 5500 1930879 1929059 1821 2001 5501 1931190 1930990 201 2002 5502 1931888 1931421 468 2003 5503 1932315 1931935 381 pir:H70638 Mycobacterium tuberculosis 38.6 58.8 114 hypothetical protein H37Rv Rv1956 2004 5504 1932879 1932373 507 2005 5505 1934358 1933522 837 2006 5506 1935912 1934971 942 sp:Y137_METJA Methanococcus jannaschii 27.1 54.6 328 hypothetical protein MJ0137 2007 5507 1936226 1936849 624 2008 5508 1937202 1937411 210 2009 5509 1938019 1937486 534 2010 5510 1938945 1940135 1191 2011 5511 1939064 1938531 534 2012 5512 1940257 1940844 588 2013 5513 1941107 1941550 444 2014 5514 1942484 1941732 753 2015 5515 1942510 1942812 303 2016 5516 1943095 1943310 216 2017 5517 1943345 1943653 309 2018 5518 1943680 1944564 885 2019 5519 1945435 1944608 828 prf:2509434A Enterococcus faecalis esp 23.0 44.1 304 surface protein 2020 5520 1945891 1945595 297 2021 5521 1946332 1945952 381 2022 5522 1947037 1946609 429 2023 5523 1948650 1947070 1581 sp:CSP1_CORGL Corynebacterium glutamicum 30.7 54.4 270 major secreted protein PS1 protein (Brevibacterium flavum) ATCC precursor 17965 csp1 2024 5524 1951450 1949021 2430 2025 5525 1952485 1951619 867 2026 5526 1954822 1952546 2277 sp:TOP3_ECOLI Escherichia coli topB 23.8 50.9 597 DNA topoisomerase III 2027 5527 1958287 1956203 2085 2028 5528 1959340 1958450 891 2029 5529 1960196 1959765 432 2030 5530 1961114 1960371 744 2031 5531 1963000 1961114 1887 sp:CSP1_CORGL Corynebacterium glutamicum 29.7 54.7 344 major secreted protein PS1 protein (Brevibacterium flavum) ATCC precursor 17965 csp1 2032 5532 1963429 1963139 291 2033 5533 1964743 1963514 1230 2034 5534 1965902 1964727 1176 2035 5535 1966267 1965911 357 2036 5536 1966301 1966984 684 sp:NUC_STAAU Staphylococcus aureus nuc 30.4 57.7 227 thermonuclease 2037 5537 1967435 1967289 147 2038 5538 1967604 1968167 564 2039 5539 1968264 1969715 1452 2040 5540 1969745 1970203 459 2041 5541 1970254 1971474 1221 2042 5542 1971672 1973090 1419 2043 5543 1973147 1973737 591 2044 5544 1973809 1974204 396 2045 5545 1974267 1974503 237 2046 5546 1975171 1975794 624 prf:2313347B Shewanella sp. ssb 24.9 59.1 225 single stranded DNA-binding protein 2047 5547 1975916 1976494 579 2048 5548 1976522 1976983 462 2049 5549 1977043 1977549 507 2050 5550 1977742 1978329 588 2051 5551 1978389 1978721 333 2052 5552 1978660 1979217 558 2053 5553 1979239 1979808 570 2054 5554 1979974 1980885 912 sp:S24D_ANOGA Anopheles gambiae AgSP24D 25.7 52.6 249 serine protease 2055 5555 1980965 1981657 693 2056 5556 1981663 1982028 366 2057 5557 1982071 1982817 747 2058 5558 1982091 1981912 180 2059 5559 1983186 1983548 363 2060 5560 1983611 1983883 273 2061 5561 1983918 1984181 264 2062 5562 1984217 1984450 234 2063 5563 1984387 1984728 342 2064 5564 1985092 1985364 273 2065 5565 1985373 1985071 303 2066 5566 1986590 1985442 1149 sp:VINT_BPML5 Mycobacterium phage L5 int 29.6 55.9 406 integrase 2067 5567 1987896 1987507 390 gsp:R23011 Brevibacterium lactofermentum 83.9 94.4 124 transposase (divided) CGL2005 ISaB1 2068 5568 1988303 1987887 417 gsp:R23011 Brevibacterium lactofermentum 70.9 84.6 117 transposase (divided) CGL2005 ISaB1 2069 5569 1988383 1988589 207 2070 5570 1988483 1988370 114 gsp:R21601 Brevibacterium lactofermentum 80.7 96.8 31 transposition repressor CGL2005 ISaB1 2071 5571 1988664 1988530 135 pir:S60889 Corynebacterium glutamicum 74.4 88.4 43 insertion element (IS3 related) orf1 2072 5572 1989605 1988778 828 gp:SCJ11_12 Streptomyces coelicolor A3(2) 31.1 53.7 270 transposase SCJ11.12 2073 5573 1990667 1991020 354 2074 5574 1990764 1989874 891 2075 5575 1991620 1991189 432 2076 5576 1992538 1991795 744 2077 5577 1994121 1992538 1584 sp:CSP1_CORGL Corynebacterium glutamicum 25.0 37.0 153 major secreted protein PS1 protein (Brevibacterium flavum) ATCC precursor 17965 csp1 2078 5578 1995294 1994608 687 sp:VINT_BPML5 Mycobacterium phage L5 int 28.7 56.1 223 integrase 2079 5579 1996088 1995783 306 pir:F64546 Helicobacter pylori 26695 39.8 76.1 88 sodium-dependent transporter HP0214 2080 5580 1996106 1996537 432 sp:YXAA_BACSU Bacillus subtilis yxaA 48.9 81.5 92 hypothetical protein 2081 5581 1996768 1997112 345 2082 5582 1997168 1997503 336 2083 5583 1997545 1998240 696 pir:C70968 Mycobacterium tuberculosis 33.5 64.4 233 riboflavin biosynthesis protein H37Rv Rv2671 ribD 2084 5584 1998289 1999542 1254 pir:E70968 Mycobacterium tuberculosis 42.5 71.9 384 potential membrane protein H37Rv Rv2673 2085 5585 1999542 1999949 408 gp:AF128264_2 Streptococcus gordonii msrA 41.3 67.5 126 methionine sulfoxide reductase 2086 5586 2000132 1999707 426 2087 5587 2001216 2000521 696 pir:H70968 Mycobacterium tuberculosis 55.2 77.2 232 hypothetical protein H37Rv Rv2676c 2088 5588 2001489 2002112 624 pir:C70528 Mycobacterium tuberculosis 55.7 78.6 201 hypothetical protein H37Rv Rv2680 2089 5589 2002072 2003334 1263 sp:RND_HAEIN Haemophilus influenzae Rd 25.9 52.8 371 ribonuclease D KW20 HI0390 rnd 2090 5590 2005309 2003402 1908 gp:AB026631_1 Streptomyces sp. CL190 dxs 55.3 78.5 618 1-deoxy-D-xylulose-5-phosphate synthase 2091 5591 2006697 2005462 1236 pir:E72298 Thermotoga maritima MSB8 25.4 52.3 472 RNA methyltransferase TM1094 2092 5592 2006698 2006979 282 2093 5593 2007637 2006777 861 pir:C70530 Mycobacterium tuberculosis 38.1 62.7 268 hypothetical protein H37Rv Rv2696c 2094 5594 2008184 2007738 447 sp:DUT_STRCO Streptomyces coelicolor A3(2) 55.0 82.1 140 deoxyuridine 5′-triphosphate SC2E9.09 dut nucleotidohydrolase 2095 5595 2008250 2008798 549 pir:E70530 Mycobacterium tuberculosis 46.0 70.7 150 hypothetical protein H37Rv Rv2698 2096 5596 2009082 2008876 207 2097 5597 2009570 2009280 291 pir:F70530 Mycobacterium tuberculosis 58.0 81.0 100 hypothetical protein H37Rv Rv2699c 2098 5598 2010539 2009724 816 sp:SUHB_ECOLI Escherichia coli k12 suhB 38.4 68.2 198 extragenic suppressor protein 2099 5599 2010555 2011382 828 sp:PPGK_MYCTU Mycobacterium tuberculosis 54.4 80.2 248 polyphosphate glucokinase H37Rv RV2702 ppgK 2100 5600 2011863 2013356 1494 prf:2204286A Corynebacterium glutamicum 98.0 98.6 500 sigma factor or RNA polymerase sigA transcription factor 2101 5601 2015496 2014162 1335 sp:YRKO_BACSU Bacillus subtilis yrkO 23.9 51.4 422 hypothetical membrane protein 2102 5602 2016121 2015585 537 2103 5603 2017966 2016257 1710 sp:Y065_MYCTU Mycobacterium tuberculosis 61.3 80.8 578 hypothetical protein H37Rv Rv2917 2104 5604 2018119 2018754 636 pir:H70531 Mycobacterium tuberculosis 32.3 59.1 127 hypothetical membrane protein H37Rv Rv2709 2105 5605 2018202 2017966 237 pir:G70531 Mycobacterium tuberculosis 65.8 85.5 76 hypothetical protein H37Rv Rv2708c 2106 5606 2018744 2020276 1533 gp:SCH5_8 Streptomyces coelicolor A3(2) 33.5 61.2 523 transferase SCH5.08c 2107 5607 2020293 2020724 432 prf:2204286C Corynebacterium glutamicum 97.2 100.0 144 hypothetical protein ATCC 13869 ORF1 2108 5608 2022266 2022949 684 pir:I40339 Corynebacterium glutamicum 98.7 99.6 228 iron dependent repressor or ATCC 13869 dtxR diphtheria toxin repressor 2109 5609 2022546 2022313 234 GP:AF010134_1 Streptomyces aureofaciens 62.0 64.0 77 putative sporulation protein 2110 5610 2022959 2023945 987 sp:GALE_BRELA Corynebacterium glutamicum 99.1 99.1 329 UDP-glucose 4-epimerase ATCC 13869 (Brevibacterium lactofermentum) galE 2111 5611 2025270 2023948 1323 2112 5612 2025423 2026379 957 pir:E70532 Mycobacterium tuberculosis 45.3 79.0 305 hypothetical protein H37Rv Rv2714 2113 5613 2026494 2029043 2550 sp:MTR4_YEAST Saccharomyces cerevisiae 24.4 50.7 661 ATP-dependent RNA helicase YJL050W dob1 2114 5614 2029177 2030157 981 sp:OXYR_ECOLI Escherichia coli oxyR 35.8 65.6 299 hydrogen peroxide-inducible genes activator 2115 5615 2031365 2030277 1089 2116 5616 2031478 2035383 3906 sp:HRPA_ECOLI Escherichia coli hrpA 49.2 76.2 1298 ATP-dependent helicase 2117 5617 2035880 2035431 450 gp:SCAJ4870_3 Streptomyces clavuligerus nrdR 61.4 86.2 145 regulatory protein 2118 5618 2036409 2035990 420 2119 5619 2036812 2037507 696 sp:LEXA_BACSU Bacillus subtilis dinR 46.9 71.6 222 SOS regulatory protein 2120 5620 2037815 2038591 777 sp:GATR_ECOLI Escherichia coli K12 gatR 33.9 67.8 245 galactitol utilization operon repressor 2121 5621 2038591 2039550 960 gp:SCE22_14 Streptomyces coelicolor A3(2) 27.2 55.6 320 phosphofructokinase (fructose 1- SCE22.14c phosphate kinase) 2122 5622 2041321 2039618 1704 sp:PT1_BACST Bacillus stearothermophilus ptsl 34.3 64.0 592 phosphoenolpyruvate-protein phosphotransferase 2123 5623 2041728 2042519 792 sp:GLPR_ECOLI Escherichia coli K12 glpR 26.7 62.6 262 glycerol-3-phosphate regulon repressor 2124 5624 2042519 2043508 990 sp:K1PF_RHOCA Rhodobacter capsulatus fruK 33.0 55.7 345 1-phosphofructokinase or 6- phosphofructokinase 2125 5625 2043736 2045571 1836 sp:PTFB_ECOLI Escherichia coli K12 fruA 43.0 69.6 549 PTS system, fructose-specific IIBC component 2126 5626 2045762 2046028 267 sp:PTHP_BACST Bacillus stearothermophilus XL- 37.0 71.6 81 phosphocarrier protein 65-6 ptsH 2127 5627 2047295 2046714 582 2128 5628 2048606 2047320 1287 sp:PYRP_BACCL Bacillus caldolyticus pyrP 39.1 70.5 407 uracil permease 2129 5629 2050107 2048650 1458 gp:AF145049_8 Streptomyces fradiae orf11* 54.4 80.0 419 ATP/GTP-binding protein 2130 5630 2050321 2051106 786 2131 5631 2051306 2051842 537 2132 5632 2052675 2051845 831 sp:DAPF_HAEIN Haemophilus influenzae Rd 33.5 64.7 269 diaminopimelate epimerase KW20 HI0750 dapF 2133 5633 2053586 2052684 903 sp:MIAA_ECOLI Escherichia coli K12 miaA 40.0 68.7 300 tRNA delta-2- isopentenylpyrophosphate transferase 2134 5634 2054283 2053609 675 2135 5635 2054403 2055761 1359 pir:B70506 Mycobacterium tuberculosis 48.5 75.7 445 hypothetical protein H37Rv Rv2731 2136 5636 2055743 2054724 1020 2137 5637 2055765 2056787 1023 2138 5638 2057788 2057120 669 pir:C70506 Mycobacterium tuberculosis 29.0 63.7 190 hypothetical membrane protein H37Rv Rv2732c 2139 5639 2059420 2057855 1566 sp:Y195_MYCLE Mycobacterium leprae 68.4 86.4 494 hypothetical protein B2235_C2_195 2140 5640 2059774 2060499 726 sp:GLUA_CORGL Corynebacterium glutamicum 99.6 99.6 242 glutamate transport ATP-binding ATCC 13032 gluA protein 2141 5641 2060414 2060196 219 GSP:Y75358 Neisseria gonorrhoeae 66.0 73.0 71 Neisserial polypeptides predicted to be useful antigens for vaccines and diagnostics 2142 5642 2061629 2062312 684 sp:GLUC_CORGL Corynebacterium glutamicum 100.0 100.0 225 glutamate transport system ATCC 13032 gluC permease protein 2143 5643 2062441 2063259 819 sp:GLUD_CORGL Corynebacterium glutamicum 99.3 99.6 273 glutamate transport system (Brevibacterium flavum) ATCC permease protein 13032 gluD 2144 5644 2063894 2063298 597 sp:RECX_MYCLE Mycobacterium leprae recX 34.5 66.9 142 regulatory protein 2145 5645 2065627 2065394 234 pir:A70878 Mycobacterium tuberculosis 40.3 71.6 67 hypothetical protein H37Rv Rv2738c 2146 5646 2066404 2065667 738 2147 5647 2066566 2067141 576 sp:BIOY_BACSH Bacillus sphaericus bioY 33.0 61.4 197 biotin synthase 2148 5648 2067168 2067866 699 sp:POTG_ECOLI Escherichia coli k12 potG 33.2 69.5 223 putrescine transport ATP-binding protein 2149 5649 2067866 2068474 609 pir:F69742 Bacillus subtilis ybaF 24.6 58.8 228 hypothetical membrane protein 2150 5650 2068703 2069392 690 pir:B60176 Mycobacterium tuberculosis 41.7 78.5 228 hypothetical protein 2151 5651 2069383 2068556 828 sp:35KD_MYCTU Mycobacterium tuberculosis 72.5 89.6 269 hypothetical protein (35kD protein) H37Rv RV2744C 2152 5652 2069936 2069616 321 pir:H70878 Mycobacterium tuberculosis 54.2 78.3 83 regulator (DNA-binding protein) H37Rv Rv2745c 2153 5653 2070512 2069997 516 sp:CINA_STRPN Streptococcus pneumoniae R6X 41.8 68.5 165 competence damage induced cinA proteins 2154 5654 2071121 2070519 603 prf:2421334D Streptococcus pyogenes pgsA 38.8 72.5 160 phosphotidylglycerophosphate synthase 2155 5655 2071315 2071599 285 pir:T10688 Arabidopsis thaliana 24.8 52.1 117 hypothetical protein ATSP:T16I18.20 2156 5656 2071624 2071740 117 gp:AF071810_1 Streptococcus pneumoniae 60.0 70.0 30 surface protein (Peumococcal DBL5 pspA surface protein A) 2157 5657 2072066 2072878 813 2158 5658 2072905 2071799 1107 prf:2119295D Escherichia coli terC 31.0 59.8 358 tellurite resistance protein 2159 5659 2076056 2073294 2763 sp:SP3E_BACSU Bacillus subtilis 168 spoIIIE 38.0 64.6 845 stage III sporulation protein E 2160 5660 2077024 2076392 633 gp:SC4G6_14 Streptomyces coelicolor A3(2) 33.3 61.0 216 hypothetical protein SC4G6.14 2161 5661 2079275 2077122 2154 sp:YOR4_CORGL Corynebacterium glutamicum 99.1 99.4 645 hypothetical protein ATCC 13032 orf4 2162 5662 2081136 2080387 750 sp:YDAP_BRELA Corynebacterium glutamicum 99.2 99.6 250 hypothetical protein (Brevibacterium lactofermentum) ATCC 13869 orf2 2163 5663 2082115 2082813 699 2164 5664 2082368 2082105 264 2165 5665 2085190 2082932 2259 prf:2217311A Streptomyces antibioticus gpsl 65.4 85.3 742 guanosine pentaphosphate synthetase 2166 5666 2085702 2085436 267 pir:F69700 Bacillus subtilis rpsO 64.0 88.8 89 30S ribosomal protein S15 2167 5667 2086826 2085879 948 prf:2518365A Leishmania major 35.1 63.3 319 nucleoside hydrolase 2168 5668 2087941 2086919 1023 sp:RIBF_CORAM Corynebacterium 56.2 79.0 329 bifunctional protein (riboflavin kinase ammoniagenes ATCC 6872 ribF and FAD synthetase) 2169 5669 2087973 2088863 891 sp:TRUB_BACSU Bacillus subtilis 168 truB 32.7 61.7 303 tRNA pseudouridine synthase B 2170 5670 2088181 2087954 228 PIR:PC4007 Corynebacterium 65.0 73.0 47 hypothetical protein ammoniagenes 2171 5671 2089868 2089218 651 gp:SC5A7_23 Streptomyces coelicolor A3(2) 42.2 62.5 237 hypothetical protein SC5A7.23 2172 5672 2090664 2089861 804 pir:B70885 Mycobacterium tuberculosis 46.9 68.9 273 phosphoesterase H37Rv Rv2795c 2173 5673 2092055 2090751 1305 pir:G70693 Mycobacterium tuberculosis 51.0 78.8 433 DNA damaged inducible protein f H37Rv Rv2836c dinF 2174 5674 2093046 2092051 996 pir:H70693 Mycobacterium tuberculosis 36.7 70.8 308 hypothetical protein H37Rv Rv2837c 2175 5675 2093501 2093055 447 sp:RBFA_BACSU Bacillus subtilis 168 rbfA 32.4 70.4 108 ribosome-binding factor A 2176 5676 2096723 2093712 3012 sp:IF2_STIAU Stigmatella aurantiaca DW4 infB 37.7 62.9 1103 translation initiation factor IF-2 2177 5677 2097179 2096844 336 gp:SC5H4_29 Streptomyces coelicolor A3(2) 44.6 66.3 83 hypothetical protein SC5H4.29 2178 5678 2098375 2097380 996 sp:NUSA_BACSU Bacillus subtilis 168 nusA 42.3 71.0 352 n-utilization substance protein (transcriptional termination/antitermination factor) 2179 5679 2098562 2099815 1254 2180 5680 2098945 2098412 534 pir:E70588 Mycobacterium tuberculosis 34.6 65.5 165 hypothetical protein H37Rv Rv2842c 2181 5681 2100240 2101841 1602 sp:DPPE_BACSU Bacillus subtilis 168 dppE 25.3 60.9 534 peptide-binding protein 2182 5682 2102023 2102946 924 sp:DPPB_ECOLI Escherichia coli K12 dppB 37.7 69.4 337 peptidetransport system permease 2183 5683 2102975 2103973 999 prf:1709239C Bacillus subtilis spo0KC 38.4 69.2 292 oligopeptide permease 2184 5684 2103973 2105703 1731 pir:H70788 Mycobacterium tuberculosis 57.6 81.3 552 peptidetransport system ABC- H37Rv Rv3663c dppD transporter ATP-binding protein 2185 5685 2107564 2105801 1764 sp:SYP_MYCTU Mycobacterium tuberculosis 67.0 84.6 578 prolyl-tRNA synthetase H37Rv Rv2845c proS 2186 5686 2107652 2108386 735 gp:SCC30_5 Streptomyces coelicolor A3(2) 39.5 65.0 243 hypothetical protein SCC30.05 2187 5687 2109147 2108389 759 sp:BCHD_RHOSH Rhodobacter sphaeroides ATCC 32.4 60.7 37 magnesium-chelatase subunit 17023 bchD 2188 5688 2110255 2109155 1101 prf:2503462AA Heliobacillus mobilis bchl 46.5 69.6 342 magnesium-chelatase subunit 2189 5689 2111183 2110434 750 prf:2108318B Propionibacterium freudenreichii 49.0 73.8 237 uroporphyrinogen III cobA methyltransferase 2190 5690 2111238 2112659 1422 sp:YPLC_CLOPE Clostridium perfringens NCIB 41.2 68.7 488 hypothetical protein 10662 ORF2 2191 5691 2113616 2112717 900 gp:SC5H1_10 Streptomyces coelicolor A3(2) 35.1 62.3 151 hypothetical protein SC5H1.10c 2192 5692 2115761 2116774 1014 pir:A70590 Mycobacterium tuberculosis 37.6 65.7 338 hypothetical protein H37Rv Rv2854 2193 5693 2116916 2118310 1395 sp:GSHR_BURCE Burkholderia cepacia AC1100 53.0 76.6 466 glutathione reductase gor 2194 5694 2117956 2117015 942 2195 5695 2118607 2119080 474 2196 5696 2119139 2119495 357 2197 5697 2119628 2120356 729 2198 5698 2121147 2120359 789 sp:AMPM_ECOLI Escherichia coli K12 map 47.2 75.8 252 methionine aminopeptidase 2199 5699 2123161 2121296 1866 prf:2224268A Streptomyces clavuligerus pcbR 27.3 56.5 630 penicillin binding protein 2200 5700 2123848 2123219 630 prf:2518330B Corynebacterium diphtheriae 44.0 72.2 216 response regulator (two-component chrA system response regulator) 2201 5701 2124996 2123848 1149 prf:2518330A Corynebacterium diphtheriae 29.5 56.8 424 two-component system sensor chrS histidine kinase 2202 5702 2125089 2126045 957 gp:AE001863_70 Deinococcus radiodurans 24.4 58.1 360 hypothetical membrane protein DRA0279 2203 5703 2126064 2126753 690 prf:2420410P Bacillus subtilis 168 yvrO 37.3 71.1 225 ABC transporter 2204 5704 2127087 2126926 162 2205 5705 2128483 2127350 1134 sp:GCPE_ECOLI Escherichia coli K12 gcpE 44.3 73.8 359 hypothetical protein (gcpE protein) 2206 5706 2128850 2129461 612 2207 5707 2129880 2128669 1212 pir:G70886 Mycobacterium tuberculosis 43.0 73.6 405 hypothetical membrane protein H37Rv Rv2869c 2208 5708 2130306 2130950 645 GSP:Y37145 Chlamydia trachomatis 36.0 43.0 147 polypeptides can be used as vaccines against Chlamydia trachomatis 2209 5709 2131078 2129903 1176 sp:DXR_ECOLI Escherichia coli K12 dxr 22.8 42.0 312 1-deoxy-D-xylulose-5-phosphate reductoisomerase 2210 5710 2131322 2131762 441 2211 5711 2131726 2131247 480 2212 5712 2133402 2131825 1578 2213 5713 2134260 2133406 855 pir:B72334 Thermotoga maritima MSB8 37.1 75.1 245 ABC transporter ATP-binding protein TM0793 2214 5714 2135551 2134454 1098 sp:YS80_MYCTU Mycobacterium tuberculosis 66.0 78.0 356 pyruvate formate-lyase 1 activating H37Rv enzyme 2215 5715 2135884 2136141 258 pir:A70801 Mycobacterium tuberculosis 41.5 74.5 94 hypothetical membrane protein H37Rv Rv3760 2216 5716 2137089 2136235 855 sp:CDSA_PSEAE Pseudomonas aeruginosa 33.3 56.5 294 phosphatidate cytidylyltransferase ATCC 15692 cdsA 2217 5717 2137840 2137286 555 sp:RRF_BACSU Bacillus subtilis 168 frr 47.0 84.3 185 ribosome recycling factor 2218 5718 2138664 2137936 729 prf:2510355C Pseudomonas aeruginosa pyrH 28.4 43.1 109 uridylate kinase 2219 5719 2138994 2139854 861 2220 5720 2139827 2139003 825 sp:EFTS_STRCO Streptomyces coelicolor A3(2) 49.6 76.8 280 elongation factor Ts SC2E1.42tsf 2221 5721 2140886 2140071 816 pir:A69699 Bacillus subtilis rpsB 54.7 83.5 254 30S ribosomal protein S2 2222 5722 2141257 2141760 504 sp:YS91_MYCTU Mycobacterium tuberculosis 46.0 58.0 120 hypothetical protein H37Rv Rv2891 2223 5723 2142686 2141763 924 prf:2417318A Proteus mirabilis xerD 40.1 68.7 297 site-specific recombinase 2224 5724 2144066 2142885 1182 sp:YX27_MYCTU Mycobacterium tuberculosis 39.8 66.8 395 hypothetical protein H37Rv Rv2896c 2225 5725 2145586 2144066 1521 sp:YX28_MYCTU Mycobacterium tuberculosis 46.6 75.8 504 Mg(2+) chelatase family protein H37Rv Rv2897c 2226 5726 2145941 2145576 366 sp:YX29_MYCTU Mycobacterium tuberculosis 40.3 72.3 119 hypothetical protein H37Rv Rv2898c 2227 5727 2146566 2146264 303 sp:YT01_MYCTU Mycobacterium tuberculosis 68.3 96.0 101 hypothetical protein H37Rv Rv2901c 2228 5728 2147192 2146566 627 sp:RNH2_HAEIN Haemophilus influenzae Rd 42.6 69.5 190 ribonuclease HII HI1059 rnhB 2229 5729 2147231 2148022 792 2230 5730 2148046 2147261 786 prf:2514288H Streptomyces lividans TK21 32.3 61.1 285 signal peptidase sipY 2231 5731 2148231 2149166 936 prf:2510361A Staphylococcus aureus sirA 25.4 59.1 323 Fe-regulated protein 2232 5732 2149571 2149359 213 2233 5733 2149972 2149634 339 sp:RL19_BACST Bacillus stearothermophilus rplS 70.3 88.3 111 50S ribosomal protein L19 2234 5734 2150335 2150997 663 sp:THIE_BACSU Bacillus subtilis 168 thiE 28.4 60.9 225 thiamine phosphate pyrophosphorylase 2235 5735 2151039 2152118 1080 gp:SC6E10_1 Streptomyces coelicolor A3(2) 34.0 64.1 376 oxidoreductase SC6E10.01 2236 5736 2152135 2152329 195 sp:THIS_ECOLI Escherichia coli K12 thiS 37.1 74.2 62 thiamine biosynthetic enzyme thiS (thiG1) protein 2237 5737 2152334 2153113 780 sp:THIG_ECOLI Escherichia coli K12 thiG 48.2 76.9 251 thiamine biosynthetic enzyme thiG protein 2238 5738 2153058 2154191 1134 prf:2417383A Emericella nidulans cnxF 30.2 56.8 437 molybdopterin biosynthesis protein 2239 5739 2156733 2154460 2274 sp:TEX_BORPE Bordetella pertussis TOHAMA I 56.6 78.7 776 transcriptional accessory protein tex 2240 5740 2157721 2156747 975 pir:A36940 Bacillus subtilis 168 degA 27.0 65.3 334 sporulation-specific degradation regulator protein 2241 5741 2159181 2157754 1428 pir:H72105 Chiamydophila pneumonlae 45.8 78.3 456 dicarboxylase translocator CWL029 ybhl 2242 5742 2159237 2159019 219 prf:2108268A Spinacia oleracea chloroplast 40.0 80.0 65 2-oxoglutarate/malate translocator 2243 5743 2160537 2159287 1251 sp:PCAB_PSEPU Pseudomonas putida pcaB 39.1 66.3 350 3-carboxy-cis, cis-muconate cycloisomerase 2244 5744 2160670 2160768 99 2245 5745 2161503 2161111 393 2246 5746 2162196 2161507 690 2247 5747 2163014 2162196 819 sp:TRMD_ECOLI Escherichia coli K12 trmD 34.8 64.8 273 tRNA (guanine-N1)- methyltransferase 2248 5748 2163098 2163745 648 gp:SCF81_27 Streptomyces coelicolor A3(2) 30.5 57.6 210 hypothetical protein SCF81.27 2249 5749 2164260 2163748 513 sp:RIMM_MYCLE Mycobacterium leprae 52.3 72.1 172 16S rRNA processing protein MLCB250.34. rimM 2250 5750 2164390 2164737 348 pir:B71881 Helicobacter pylori J99 jhp0839 29.0 66.7 69 hpothetical protein 2251 5751 2165309 2164815 495 pir:C47154 Bacillus subtilis 168 rpsP 47.0 79.5 83 30S ribosomal protein S16 2252 5752 2165523 2166098 576 pir:T14151 Mus musculus inv 32.1 61.7 196 inversin 2253 5753 2166990 2166124 867 prf:2512328G Streptococcus agalactiae cylB 26.6 69.1 256 ABC transporter 2254 5754 2167865 2166990 876 prf:2220349C Pyrococcus horikoshii OT3 mtrA 35.5 63.8 318 ABC transporter 2255 5755 2169584 2167944 1641 sp:SR54_BACSU Bacillus subtilis 168 ffh 58.7 78.2 559 signal recognition particle protein 2256 5756 2170426 2171058 633 2257 5757 2171715 2172131 417 2258 5758 2172209 2172877 669 2259 5759 2175288 2173759 1530 sp:FTSY_ECOLI Escherichia coli K12 ftsY 37.0 66.1 505 cell division protein 2260 5760 2176046 2175888 159 2261 5761 2176402 2177103 702 2262 5762 2179502 2176110 3393 sp:AMYH_YEAST Saccharomyces cerevisiae 22.4 46.2 1144 glucan 1,4-alpha-glucosidase or S288C YIR019C sta1 glucoamylase S1/S2 precursor 2263 5763 2180918 2181880 963 2264 5764 2183092 2179628 3465 sp:Y06B_MYCTU Mycobacterium tuberculosis 48.3 72.6 1206 chromosome segregation protein H37Rv Rv2922c smc 2265 5765 2183391 2183110 282 sp:ACYP_MYCTU Mycobacterium tuberculosis 51.1 73.9 92 acylphosphatase H37Rv RV2922.1C 2266 5766 2185258 2183405 1854 2267 5767 2186208 2185351 858 sp:YFER_ECOLI Escherichia coli K12 yfeR 23.9 60.0 305 transcriptional regulator 2268 5768 2186299 2187129 831 pir:S72748 Mycobacterium leprae 39.3 73.5 257 hypothetical membrane protein MLCL581.28c 2269 5769 2187160 2187342 183 2270 5770 2187679 2187233 447 2271 5771 2188306 2187692 615 gp:DNINTREG_3 Dichelobacter nodosus gep 46.8 76.6 188 cation efflux system protein 2272 5772 2189170 2188313 858 sp:FPG_ECOLI Escherichia coli K12 mutM or 36.1 66.7 285 formamidopyrimidine-DNA fpg glycosylase 2273 5773 2189906 2189166 741 pir:B69693 Bacillus subtilis 168 rncS 40.3 76.5 221 ribonuclease III 2274 5774 2190439 2189906 534 sp:Y06F_MYCTU Mycobacterium tuberculosis 35.8 62.5 176 hypothetical protein H37Rv Rv2926c 2275 5775 2191328 2190540 789 sp:Y06G_MYCTU Mycobacterium tuberculosis 50.0 76.9 238 hypothetical protein H37Rv Rv2927c 2276 5776 2191522 2193165 1644 prf:2104260G Streptomyces verticillus 28.3 55.6 559 transport protein 2277 5777 2193165 2194694 1530 sp:CYDC_ECOLI Escherichia coli K12 cydC 26.6 58.8 541 ABC transporter 2278 5778 2196883 2198004 1122 gp:SC9C7_2 Streptomyces coelicolor A3(2) 35.3 62.6 388 hypothetical protein SC9C7.02 2279 5779 2198447 2198007 441 2280 5780 2198475 2199758 1284 pir:A72322 Thermotoga maritima MSB8 21.0 43.7 405 hypothetical protein TM0896 2281 5781 2199808 2201070 1263 sp:HIPO_CAMJE Campylobacter jejuni ATCC 32.9 64.3 353 peptidase 43431 hipO 2282 5782 2201408 2201073 336 pir:S38197 Arabidopsis thaliana SUC1 27.1 51.9 133 sucrose transport protein 2283 5783 2201584 2201450 135 2284 5784 2201869 2201594 276 2285 5785 2204541 2201992 2550 prf:2513410A Thermococcus litoralis malP 36.1 67.4 814 maltodextrin phosphorylase / glycogen phosphorylase 2286 5786 2205490 2204591 900 sp:YFIE_BACSU Bacillus subtilis 168 yfiE 33.9 66.4 295 hypothetical protein 2287 5787 2208249 2207302 948 sp:LGT_STAAU Staphylococcus aureus FDA 485 31.4 65.5 264 prolipoprotein diacylglyceryl lgt transferase 2288 5788 2209167 2208367 801 sp:TRPG_EMENI Emericella nidulans trpC 29.6 62.1 169 indole-3-glycerol-phosphate synthase/anthranilate synthase component II 2289 5789 2209888 2209232 657 pir:H70556 Mycobacterium tuberculosis 29.4 58.8 228 hypothetical membrane protein H37Rv Rv1610 2290 5790 2210273 2209920 354 sp:HIS3_RHOSH Rhodobacter sphaeroides ATCC 52.8 79.8 89 phosphoribosyl-AMP cyclohydrolase 17023 hisI 2291 5791 2211046 2210273 774 sp:HIS6_CORG Corynebacterium glutamicum 97.3 97.7 258 cyclase AS019 hisF 2292 5792 2211875 2211051 825 prf:2419176B Corynebacterium glutamicum 94.0 94.0 241 inositol monophosphate AS019 impA phosphatase 2293 5793 2212619 2211882 738 gp:AF051846_1 Corynebacterium glutamicum 95.9 97.6 245 phosphoribosylformimino-5- AS019 hisA aminoimidazole carboxamide ribotide isomerase 2294 5794 2213273 2212641 633 gp:AF060558_1 Corynebacterium glutamicum 86.7 92.4 210 glutamine amidotransferase AS019 hisH 2295 5795 2215586 2214321 1266 sp:CMLR_STRLI Streptomyces lividans 66 cmlR 25.6 54.0 402 chloramphenicol resistance protein or transmembrane transport protein 2296 5796 2215863 2215639 225 2297 5797 2216474 2215869 606 sp:HIS7_STRCO Streptomyces coelicolor A3(2) 52.5 81.8 198 imidazoleglycerol-phosphate hisB dehydratase 2298 5798 2217591 2216494 1098 sp:HIS8_STRCO Streptomyces coelicolor A3(2) 57.2 79.3 362 histidinol-phosphate hisC aminotransferase 2299 5799 2218925 2217600 1326 sp:HISX_MYCSM Mycobacterium smegmatis 63.8 85.7 439 histidinol dehydrogenase ATCC 607 hisD 2300 5800 2219159 2220358 1200 gp:SPBC215_13 Schizosaccharomyces pombe 27.2 54.4 342 serine-rich secreted protein SPBC215.13 2301 5801 2221109 2220459 651 2302 5802 2221611 2221919 309 2303 5803 2221828 2221187 642 prf:2321269A Leishmania donovani SAcP-1 29.4 59.7 211 histidine secretory acid phosphatase 2304 5804 2221958 2222518 561 pir:RPECR1 Escherichia coli plasmid RP1 28.9 60.8 204 tet repressor protein tetR 2305 5805 2222528 2225035 2508 prf:2307203B Sulfolobus acidocaldarius treX 47.4 75.5 722 glycogen debranching enzyme 2306 5806 2225149 2225949 801 pir:E70572 Mycobacterium tuberculosis 50.0 76.0 258 hypothetical protein H37Rv Rv2622 2307 5807 2226763 2225990 774 gp:SC2G5_27 Streptomyces coelicolor A3(2) 29.9 55.2 268 oxidoreductase SC2G5.27c gip 2308 5808 2227779 2226769 1011 prf:2503399A Sinorhizobium meliloti idhA 35.0 60.9 343 myo-inositol 2-dehydrogenase 2309 5809 2227906 2228901 996 sp:GALR_ECOLI Escherichia coli K12 galR 30.4 64.4 329 galactitol utilization operon repressor 2310 5810 2229896 2229099 798 sp:FHUC_BACSU Bacillus subtilis 168 fhuC 32.9 68.3 246 ferrichrome transport ATP-binding protein or ferrichrome ABC transporter 2311 5811 2230937 2229900 1038 prf:2423441E Vibrio cholerae hutC 36.8 71.1 332 hemin permease 2312 5812 2231294 2230947 348 pir:G70046 Bacillus subtilis 168 yvrC 30.1 68.0 103 iron-binding protein 2313 5813 2231932 2231339 594 pir:G70046 Bacillus subtilis 168 yvrC 34.6 67.6 182 iron-binding protein 2314 5814 2232456 2232016 441 sp:YTFH_ECOLI Escherichia coli K12 ytfH 38.1 73.5 113 hypothetical protein 2315 5815 2232928 2234070 1143 gp:SCI8_12 Streptomyces coelicolor A3(2) 23.4 50.1 355 DNA polymerase III epsilon chain SCI8.12 2316 5816 2234158 2234763 606 2317 5817 2234852 2237284 2433 pir:S65769 Arthrobacter sp. Q36 treY 42.0 68.6 814 maltooligosyl trehalose synthase 2318 5818 2237331 2238353 1023 gp:AE002006_4 Deinococcus radiodurans 27.6 52.8 322 hypothetical protein DR1631 2319 5819 2239092 2238694 399 2320 5820 2240042 2239845 198 2321 5821 2240246 2240058 189 2322 5822 2240563 2239508 1056 2323 5823 2240681 2241724 1044 sp:LXA1_PHOLU Photorhabdus luminescens 20.5 54.4 375 alkanal monooxygenase alpha chain ATCC 29999 luxA 2324 5824 2242115 2241738 378 gp:SC7H2_5 Streptomyces coelicolor A3(2) 58.3 79.2 120 hypothetical protein SC7H2.05 2325 5825 2242359 2242129 231 2326 5826 2243035 2244819 1785 pir:S65770 Arthrobacter sp. Q36 treZ 46.3 72.4 568 maltooligosyltrehalose trehalohydrolase 2327 5827 2243043 2242393 651 sp:YVYE_BACSU Bacillus subtilis 168 36.5 72.4 214 hypothetical protein 2328 5828 2246171 2244864 1308 sp:THD1_CORGL Corynebacterium glutamicum 99.3 99.3 436 threonine dehydratase ATCC 13032 ilvA 2329 5829 2246386 2246892 507 2330 5830 2246450 2246295 156 2331 5831 2248208 2247006 1203 pir:S57636 Catharanthus roseus metE 22.7 49.6 415 Corynebacterium glutamicumASO19 2332 5832 2251939 2248358 3582 prf:2508371A Streptomyces coelicolor A3(2) 53.3 80.5 1183 DNA polymerase III dnaE 2333 5833 2252017 2252856 840 sp:RARD_ECOLI Escherichia coli K12 rarD 37.6 73.8 279 chloramphenicol sensitive protein 2334 5834 2253192 2253659 468 sp:HISJ_CAMJE Campylobacter jejuni DZ72 hisJ 21.5 55.7 149 histidine-binding protein precursor 2335 5835 2253725 2254642 918 pir:D69548 Archaeoglobus fulgidus AF2388 22.7 64.7 198 hypothetical membrane protein 2336 5836 2255558 2254683 876 sp:GS39_BACSU Bacillus subtilis 168 ydaD 48.2 80.0 280 short chain dehydrogenase or general stress protein 2337 5837 2257024 2255738 1287 sp:DCDA_PSEAE Pseudomonas aeruginosa lysA 22.9 47.6 445 diaminopimelate (DAP) decarboxylase 2338 5838 2259312 2258362 951 sp:CYSM_ALCEU Alcaligenes eutrophus CH34 32.8 64.3 314 cysteine synthase cysM 2339 5839 2259999 2259421 579 2340 5840 2260931 2260002 930 sp:RLUD_ECOLI Escherichia coli K12 rluD 36.5 61.0 326 ribosomal large subunit pseudouridine synthase D 2341 5841 2261467 2260934 534 sp:LSPA_PSEFL Pseudomonas fluorescens NCIB 33.8 61.7 154 lipoprotein signal peptidase 10586 lspA 2342 5842 2261688 2262689 1002 2343 5843 2262850 2264499 1650 pir:S67863 Streptomyces antibioticus oleB 36.4 64.0 550 oleandomycin resistance protein 2344 5844 2264996 2265298 303 2345 5845 2265108 2264509 600 prf:2422382P Rhodococcus erythropolis orf17 36.7 57.6 158 hypothetical protein 2346 5846 2265420 2266394 975 sp:ASPG_BACLI Bacillus licheniformis 31.2 62.0 321 L-asparaginase 2347 5847 2268297 2266897 1401 sp:DINP_ECOLI Escherichia coli K12 dinP 31.8 60.7 371 DNA-damage-inducible protein P 2348 5848 2269245 2268388 858 sp:YBIF_ECOLI Escherichia coli K12 ybiF 31.5 61.5 286 hypothetical membrane protein 2349 5849 2270261 2269260 1002 gp:SCF51_6 Streptomyces coelicolor A3(2) 44.3 73.1 334 transcriptional regulator SCF51.06 2350 5850 2270304 2270435 132 2351 5851 2270884 2270258 627 gp:SCF51_5 Streptomyces coelicolor A3(2) 42.0 67.0 212 hypothetical protein SCF51.05 2352 5852 2274149 2270988 3162 sp:SYIC_YEAST Saccharomyces cerevisiae 38.5 65.4 1066 isoleucyl-tRNA synthetase A364A YBL076C ILS1 2353 5853 2274688 2274473 216 2354 5854 2275861 2274767 1095 2355 5855 2276637 2276353 285 pir:F70578 Mycobacterium tuberculosis 46.3 73.2 82 hypothetical membrane protein H37Rv Rv2146c 2356 5856 2277336 2276881 456 gp:BLFTSZ_6 Brevibacterium lactofermentum 99.3 99.3 152 hypothetical protein (putative YAK 1 orf6 protein) 2357 5857 2278078 2277416 663 sp:YFZ1_CORGL Corynebacterium glutamicum 97.7 99.6 221 hypothetical protein 2358 5858 2278859 2278122 738 prf:2420425C Brevibacterium lactofermentum 99.2 100.0 246 hypothetical protein yfih 2359 5859 2279155 2279640 486 GP:AB028868_1 Mus musculus P4(21)n 39.0 51.0 117 hypothetical protein 2360 5860 2280215 2278890 1326 sp:FTSZ_BRELA Brevibacterium lactofermentum 98.6 98.6 442 cell division protein ftsZ 2361 5861 2281135 2280470 666 gsp:W70502 Corynebacterium glutamicum 99.6 100.0 222 cell division initiation protein or cell ftsQ division protein 2362 5862 2282623 2281166 1458 gp:AB015023_1 Corynebacterium glutamicum 99.4 99.8 486 UDP-N-acetylmuramate-alanine murC ligase 2363 5863 2283776 2282661 1116 gp:BLA242646_3 Brevibacterium lactofermentum 98.9 99.5 372 UDP-N-acetylglucosamine-N- ATCC 13869 murG acetylmuramyl-(pentapeptide) pyrophosphoryl-undecaprenol N- acetylglucosamine pyrophosphoryl- undecaprenol N-acetylglucosamine 2364 5864 2285431 2283782 1650 gp:BLA242646_2 Brevibacterium lactofermentum 99.4 99.6 490 cell division protein ATCC 13869 ftsW 2365 5865 2285904 2285437 468 gp:BLA242646_1 Brevibacterium lactofermentum 99.1 99.1 110 UDP-N-acetylmuramoylalanine-D- ATCC 13869 murD glutamate ligase 2366 5866 2286272 2286655 384 2367 5867 2286499 2286831 333 2368 5868 2287959 2286862 1098 sp:MRAY_ECOLI Escherichia coli K12 mraY 38.6 63.8 365 phospho-n-acetylmuramoyl- pentapeptide 2369 5869 2289510 2287969 1542 sp:MURF_ECOLI Escherichia coli K12 murF 35.0 64.2 494 UDP-N-acetylmuramoylalanyl-D- glutamyl-2,6-diaminopimelate-D- alanyl-D-alanyl ligase 2370 5870 2291073 2289523 1551 sp:MURE_BACSU Bacillus subtilis 168 murE 37.7 67.6 491 UDP-N-acetylmuramoylalanyl-D- glutamyl-2,6-diaminopimelate-D- alanyl-D-alanyl ligase 2371 5871 2291197 2290973 225 GSP:Y33117 Brevibacterium lactofermentum 100.0 100.0 57 penicillin binding protein ORF2 pbp 2372 5872 2293164 2291212 1953 pir:S54872 Pseudomonas aeruginosa pbpB 28.2 58.8 650 penicillin-binding protein 2373 5873 2294117 2293323 795 2374 5874 2295127 2294117 1011 pir:A70581 Mycobacterium tuberculosis 55.1 79.3 323 hypothetical protein H37Rv Rv2165c 2375 5875 2295804 2295376 429 gp:MLCB268_11 Mycobacterium leprae 72.0 88.8 143 hypothetical membrane protein MLCB268.11c 2376 5876 2296898 2296512 387 pir:C70935 Mycobacterium tuberculosis 39.4 69.3 137 hypothetical protein H37Rv Rv2169c 2377 5877 2297653 2297231 423 2378 5878 2297866 2298438 573 gp:MLCB268_13 Mycobacterium leprae 36.3 65.3 190 hypothetical protein MLCB268.13 2379 5879 2299428 2298451 978 sp:METF_STRLI Streptomyces lividans 1326 42.6 70.6 303 5,10-methylenetetrahydrofolate metF reductase 2380 5880 2299524 2300636 1113 pir:S32168 Myxococcus xanthus DK1050 30.1 62.0 329 dimethylallyltranstransferase ORF1 2381 5881 2300706 2302175 1470 gp:MLCB268_16 Mycobacterium leprae 35.7 69.6 484 hypothetical membrane protein MLCB268.17 2382 5882 2302179 2302685 507 2383 5883 2302619 2302251 369 pir:A70936 Mycobacterium tuberculosis 43.2 68.8 125 hypothetical protein H37Rv Rv2175c 2384 5884 2302833 2304980 2148 gp:AB019394_1 Streptomyces coelicolor A3(2) 34.2 62.4 684 eukaryotic-type protain kinase pkaF 2385 5885 2303690 2303040 651 2386 5886 2304983 2306218 1236 gp:MLCB268_21 Mycobacterium leprae 30.7 58.4 411 hypothetical membrane protein MLCB268.23 2387 5887 2306314 2307621 1308 pir:G70936 Mycobacterium tuberculosis 30.4 62.0 434 hypothetical membrane protein H37Rv Rv2181 2388 5888 2309082 2307697 1386 gp:AF260581_2 Amycolatopsis mediterranei 66.9 87.9 462 3-deoxy-D-arabino-heptulosonate-7- phosphate synthase 2389 5889 2309676 2309173 504 gp:MLCB268_20 Mycobacterium leprae 58.4 77.7 166 hypothetical protein MLCB268 21c 2390 5890 2309835 2312252 2418 pir:G70936 Mycobacterium tuberculosis 35.1 64.5 428 hypothetical membrane protein H37Rv Rv2181 2391 5891 2312360 2313808 1449 sp:CSP1_CORGL Corynebacterium glutamicum 28.2 57.1 440 major secreted protein PS1 protein (Brevibacterium flavum) ATCC precursor 17965 csp1 2392 5892 2313833 2314036 204 2393 5893 2314092 2313916 177 2394 5894 2315423 2314236 1188 gp:AF096280_3 Corynebacterium glutamicum 100.0 100.0 249 hypothetical membrane protein ATCC 13032 2395 5895 2316412 2315678 735 gp:AF096280_2 Corynebacterium glutamicum 100.0 100.0 245 acyltransferase ATCC 13032 2396 5896 2318775 2317633 1143 gp:SC6G10_5 Streptomyces coelicolor A3(2) 50.1 75.7 383 glycosyl transferase SC6G10.05c 2397 5897 2319850 2318804 1047 sp:P60_LISIV Listeria ivanovii iap 26.4 60.8 296 protein P60 precursor (invasion- associated-protein) 2398 5898 2320594 2319968 627 sp:P60_LISGR Listeria grayi iap 33.0 61.3 191 protein P60 precursor (invasion- associated-protein) 2399 5899 2323073 2321472 1602 prf:2503462K Heliobacillus mobilis petB 34.3 64.7 201 ubiquinol-cytochrome c reductase cytochrome b subunit 2400 5900 2323759 2323088 672 gp:AF107888_1 Streptomyces lividans qcrA 37.9 57.1 203 ubiquinol-cytochrome c reductase iron-sulfur subunit (Rieske [eFe-2S] iron-sulfur protein cyoB 2401 5901 2325195 2324311 885 sp:Y005_MYCTU Mycobacterium tuberculosis 58.6 83.1 278 ubiquinol-cytochrome c reductase H37Rv Rv2194 qcrC cytochrome c 2402 5902 2325887 2325273 615 sp:COX3_SYNVU Synechococcus vulcanus 36.7 70.7 188 cytochrome c oxidase subunit III 2403 5903 2326273 2326121 153 2404 5904 2326900 2326472 429 sp:Y00A_MYCTU Mycobacterium tuberculosis 38.6 71.0 145 hypothetical membrane protein H37Rv Rv2199c 2405 5905 2327997 2326921 1077 sp:COX2_RHOSH Rhodobacter sphaeroides ctaC 28.7 53.9 317 cytochrome c oxidase subunit II 2406 5906 2328516 2330435 1920 gp:AB029550_1 Corynebacterium glutamicum 99.7 99.8 640 glutamine-dependent KY9611 ltsA amidotransferase or asparagine synthetase (lysozyme insensitivity protein) 2407 5907 2330927 2330586 342 gp:AB029550_2 Corynebacterium glutamicum 100.0 100.0 114 hypothetical protein KY9611 orf1 2408 5908 2331200 2331967 768 gp:MLCB22_2 Mycobacterium leprae 35.0 60.2 246 hypothetical membrane protein MLCB22.07 2409 5909 2331974 2332495 522 pir:S52220 Rhodobacter capsulatus cobP 43.0 64.0 172 cobinamide kinase 2410 5910 2332512 2333600 1089 sp:COBU_PSEDE Pseudomonas denitrificans 37.8 66.9 341 nicotinate-nucleotide- cobU dimethylbenzimidazole phosphoribosyltransferase 2411 5911 2333615 2334535 921 sp:COBV_PSEDE Pseudomonas denitrificans cobV 25.3 49.8 305 cobalamin (5′-phosphate) synthase 2412 5912 2334717 2334481 237 2413 5913 2335741 2335028 714 prf:2414335A Streptomyces clavuligerus car 38.6 68.5 241 clavulanate-9-aldehyde reductase 2414 5914 2337051 2335915 1137 sp:ILVE_MYCTU Mus musculus BCAT1 40.1 70.3 364 branched-chain amino acid aminotransferase 2415 5915 2337235 2338734 1500 gp:PPU010261_1 Pseudomonas putida ATCC 36.3 65.9 493 leucyl aminopeptidase 12633 pepA 2416 5916 2339140 2338748 393 prf:2110282A Saccharopolyspora erythraea 40.2 67.0 97 hypothetical protein ORF1 2417 5917 2339269 2341293 2025 gp:AF047034_2 Streptomyces seoulensis pdhB 48.9 68.5 691 dihydrolipoamide acetyltransferase 2418 5918 2340804 2339440 1365 2419 5919 2341412 2342164 753 gp:AB020975_1 Arabidopsis thaliana 36.7 65.7 210 lipoyltransferase 2420 5920 2342304 2343347 1044 sp:LIPA_PELCA Pelobacter carbinolicus GRA BD 44.6 70.9 285 lipoic acid synthetase 1 lipA 2421 5921 2343479 2344258 780 sp:Y00U_MYCTU Mycobacterium tuberculosis 45.5 76.7 257 hypothetical membrane protein H37Rv Rv2219 2422 5922 2344431 2346047 1617 sp:YIDE_ECOLI Escherichia coli K12 yidE 32.9 67.8 559 hypothetical membrane protein 2423 5923 2347491 2346289 1203 gp:AF189147_1 Corynebacterium glutamicum 100.0 100.0 401 transposase (ISCg2) ATCC 13032 tnp 2424 5924 2347505 2347804 300 2425 5925 2348548 2348078 471 gp:SCSF7_34 Streptomyces coelicolor A3(2) 41.4 63.7 157 hypothetical membrane protein SC5F7.04c 2426 5926 2350620 2350408 213 2427 5927 2351022 2351996 975 31.0 44.0 145 mutator mutT domain protein 2428 5928 2351310 2350912 399 pir:B72308 Thermotoga maritima MSB8 36.7 65.6 128 hypothetical protein TM1010 2429 5929 2351909 2351310 600 2430 5930 2351980 2352828 849 sp:LUXA_VIBHA Vibrio harveyi luxA 25.0 60.9 220 alkanal monooxygenase alpha chain (bacterial luciferase alpha chain) 2431 5931 2352833 2353225 393 pir:A72404 Thermotoga maritima MSB8 40.5 73.0 111 protein synthesis inhibitor TM0215 (translation initiation inhibitor) 2432 5932 2355156 2355398 243 2433 5933 2355440 2355180 261 2434 5934 2355521 2356843 1323 prf:2203345H Escherichia coli hpaX 21.9 53.4 433 4-hydroxyphenylacetate permease 2435 5935 2356794 2357354 561 gp:SCGD3_10 Streptomyces coelicolor A3(2) 42.4 72.8 158 transmembrane transport protein SCGD3.10c 2436 5936 2357264 2357707 444 gp:SCGD3_10 Streptomyces coelicolor A3(2) 31.4 66.1 118 transmembrane transport protein SCGD3.10c 2437 5937 2357484 2357290 195 2438 5938 2357726 2358130 405 2439 5939 2358695 2358153 543 2440 5940 2359416 2358772 645 sp:HMUO_CORDI Corynebacterium diphtheriae C7 57.9 78.0 214 heme oxygenase hmuO 2441 5941 2362748 2359614 3135 gp:SCY17736_4 Streptomyces coelicolor A3(2) 43.4 67.0 809 glutamate-ammonia-ligase glnE adenylytransferase 2442 5942 2364155 2362818 1338 sp:GLNA_THEMA Thermotoga maritima MSB8 43.5 73.0 441 glutamine synthetase glnA 2443 5943 2364352 2365455 1104 gp:SCE9_39 Streptomyces coelicolor A3(2) 26.8 54.1 392 hypothetical protein SCE9.39c 2444 5944 2365587 2367413 1827 sp:Y017_MYCTU Mycobacterium tuberculosis 33.4 58.2 601 hypothetical protein H37Rv Rv2226 2445 5945 2367652 2367473 180 gp:SCC75A_11 Streptomyces coelicolor A3(2) 38.9 55.6 54 hypothetical protein SCC75A.11c. 2446 5946 2367791 2369083 1293 sp:GAL1_HUMAN Homo sapiens galK1 24.9 53.7 374 galactokinase 2447 5947 2370381 2369116 1266 gp:AF174645_1 Brucella abortus vacB 27.1 54.5 358 virulence-associated protein 2448 5948 2370423 2370908 486 2449 5949 2372557 2371412 1146 sp:Y019_MYCTU Mycobacterium tuberculosis 54.7 75.1 382 bifunctional protein (ribonuclease H H37Rv Rv2228c and phosphoglycerate mutase) 2450 5950 2372561 2373289 729 2451 5951 2373289 2372573 717 sp:Y01A_MYCTU Mycobacterium tuberculosis 26.5 58.6 249 hypothetical protein H37Rv Rv2229c 2452 5952 2374462 2373323 1140 sp:Y01B_MYCTU Mycobacterium tuberculosis 49.2 76.2 378 hypothetical protein H37Rv Rv2230c 2453 5953 2374544 2375197 654 sp:GPH_ECOLI Escherichia coli K12 gph 26.0 54.4 204 phosphoglycolate phosphatase 2454 5954 2375214 2375684 471 sp:PTPA_STRCO Streptomyces coelicolor A3(2) 46.2 63.5 156 low molecular weight protein- SCQ11.04c ptpA tyrosine-phosphatase 2455 5955 2375767 2376720 954 sp:Y01G_MYCTU Mycobacterium tuberculosis 40.9 65.5 281 hypothetical protein H37Rv Rv2235 2456 5956 2377390 2376998 393 sp:YI21_BURCE Burkholderia cepacia 32.6 56.6 129 insertion element (IS402) 2457 5957 2377726 2377484 243 2458 5958 2377899 2378276 378 gp:SC8F4_22 Streptomyces coelicolor A3(2) 30.4 57.8 135 transcriptional regulator SC8F4.22c 2459 5959 2378292 2378489 198 2460 5960 2379312 2378884 429 sp:Y01K_MYCTU Mycobacterium tuberculosis 55.2 77.6 134 hypothetical protein H37Rv Rv2239c 2461 5961 2379426 2379770 345 2462 5962 2380033 2382744 2712 gp:AF047034_4 Streptomyces seoulensis pdhA 55.9 78.9 910 pyruvate dehydrogenase component 2463 5963 2382240 2380765 1476 2464 5964 2383615 2382827 789 sp:GLNQ_ECOLI Escherichia coli K12 glnQ 33.7 62.8 261 ABC transporter or glutamine transport ATP-binding protein 2465 5965 2384464 2385426 963 2466 5966 2384509 2383622 888 sp:RBSC_BACSU Bacillus subtilis 168 rbsC 25.4 58.7 283 ribose transport system permease protein 2467 5967 2385447 2384509 939 pir:H71693 Rickettsia prowazekii Madrid E 26.2 62.9 286 hypothetical protein RP367 2468 5968 2385771 2386580 810 sp:CBPA_DICDI Dictyostelium discoideum AX2 41.6 55.2 125 calcium binding protein cbpA 2469 5969 2386284 2385913 372 2470 5970 2387627 2386614 1014 gp:SC6G4_24 Streptomyces coelicolor A3(2) 29.6 55.7 352 lipase or hydrolase SC6G4.24 2471 5971 2387667 2387957 291 sp:ACP_MYXXA Myxococcus xanthus ATCC 42.7 80.0 75 acyl carier protein 25232 acpP 2472 5972 2387997 2388821 825 sp:NAGD_ECOLI Escherichia coli K12 nagD 43.9 75.5 253 N-acetylglucosamine-6-phosphate deacetylase 2473 5973 2388838 2389869 1032 gp:AE001968_4 Deinococcus radiodurans 33.6 65.7 289 hypothetical protein DR1192 2474 5974 2390904 2390434 471 2475 5975 2392008 2391184 825 gp:SC4A7_8 Streptomyces coelicolor A3(2) 52.4 75.3 271 hypothetical protein SC4A7.08 2476 5976 2392566 2392075 492 2477 5977 2393349 2392579 771 2478 5978 2393425 2393970 546 2479 5979 2394437 2393973 465 2480 5980 2394594 2394935 342 2481 5981 2395204 2396763 1560 sp:PPBD_BACSU Bacillus subtilis 168 phoD 34.2 64.7 530 alkaline phosphatase D precursor 2482 5982 2395986 2395273 714 2483 5983 2397264 2399099 1836 gp:SCI51_17 Streptomyces coelicolor A3(2) 44.4 73.1 594 hypothetical protein SCI51.17 2484 5984 2399158 2399397 240 pir:G70661 Mycobacterium tuberculosis 41.2 72.1 68 hypothetical protein H37Rv Rv2342 2485 5985 2400342 2399668 675 2486 5986 2401303 2399405 1899 prf:2413330B Mycobacterium smegmatis 59.1 82.9 633 DNA primase dnaG 2487 5987 2401373 2401834 462 gp:XXU39467_1 Streptomyces aureofaciens BMK 49.0 67.4 98 ribonuclease Sa 2488 5988 2401838 2402080 243 2489 5989 2403165 2402530 636 2490 5990 2404012 2402144 1869 gp:AF058788_1 Mycobacterium smegmatis 59.1 82.2 636 L-glutamine:D-fructose-6-phosphate mc2155 glmS amidotransferase 2491 5991 2404523 2404846 324 2492 5992 2405671 2406822 1152 2493 5993 2406258 2404987 1272 prf:2413330A Mycobacterium smegmatis dgt 54.6 76.3 414 deoxyguanosinetriphosphate triphosphohydrolase 2494 5994 2406936 2406262 675 gp:NMA1Z2491_235 Neisseria meningitidis NMA0251 30.4 59.7 171 hypothetical protein 2495 5995 2406993 2409029 2037 pir:B70662 Mycobacterium tuberculosis 31.1 63.6 692 hypothetical protein H37Rv Rv2345 2496 5996 2410264 2409779 486 gp:AE003565_26 Drosophila melanogaster 24.6 54.4 138 hypothetical protein CG10592 2497 5997 2410861 2410280 582 2498 5998 2412338 2410956 1383 pir:S58522 Thermus aquaticus HB8 46.1 69.9 508 glycyl-tRNA synthetase 2499 5999 2412580 2412948 369 pir:E70585 Mycobacterium tuberculosis 49.4 73.0 89 bacterial regulatory protein, arsR H37Rv Rv2358 furB family 2500 6000 2412992 2413423 432 sp:FUR_ECOLI Escherichia coli K12 fur 34.9 70.5 132 ferric uptake regulation protein 2501 6001 2413568 2415118 1551 pir:A70539 Mycobacterium tuberculosis 24.8 46.7 529 hypothetical protein (conserved in H37Rv Rv1128c C.glutamicum?) 2502 6002 2416089 2415298 792 gp:AF162938_1 Streptomyces coelicolor A3(2) 40.6 67.0 224 hypothetical membrane protein h3u 2503 6003 2417099 2416371 729 sp:UPPS_MICLU Micrococcus luteus B-P 26 uppS 43.4 71.2 233 undecaprenyl diphosphate synthase 2504 6004 2417947 2417222 726 pir:A70586 Mycobacterium tuberculosis 45.7 74.3 245 hypothetical protein H37Rv Rv2362c 2505 6005 2418883 2417969 915 gp:AF072811_1 Streptococcus pneumoniae era 39.5 70.3 296 Era-like GTP-binding protein 2506 6006 2420309 2418990 1320 sp:Y1DE_MYCTU Mycobacterium tuberculosis 52.8 82.4 432 hypothetical membrane protein H37Rv Rv2366 2507 6007 2420900 2420313 588 sp:YN67_MYCTU Mycobacterium tuberculosis 65.0 86.0 157 hypothetical protein H37Rv Rv2367c 2508 6008 2420973 2421236 264 GSP:Y75650 Neisseria meningitidis 45.0 50.0 85 Neisserial polypeptides predicted to be useful antigens for vaccines and diagnostics 2509 6009 2421949 2420900 1050 sp:PHOL_MYCTU Mycobacterium tuberculosis 61.1 84.6 344 phosphate starvation inducible H37Rv Rv2368c phoH protein 2510 6010 2422697 2421975 723 gp:SCC77_19 Streptomyces coelicolor A3(2) 44.0 75.4 248 hypothetical protein SCC77.19c. 2511 6011 2422850 2423791 942 2512 6012 2423845 2422700 1146 prf:2421342B Streptomyces albus dnaJ2 47.1 77.4 380 heat shock protein dnaJ 2513 6013 2424937 2423915 1023 prf:2421342A Streptomyces albus hrcA 48.2 79.6 334 heat-inducible transcriptional repressor (groEL repressor) 2514 6014 2425954 2424965 990 prf:2318256A Bacillus stearothermophilus 33.1 64.1 320 oxygen-independent hemN coproporphyrinogen III oxidase 2515 6015 2426181 2426699 519 sp:AGA1_YEAST Saccharomyces cerevisiae 36.6 64.9 134 agglutinin attachment subunit YNR044WAGA1 precursor 2516 6016 2427468 2426776 693 2517 6017 2428184 2427807 378 2518 6018 2430028 2428184 1845 gp:SC6G10_4 Streptomyces coelicolor A3(2) 48.0 75.1 611 long-chain-fatty-acid-CoA ligase SC6G10.04 2519 6019 2430296 2432413 2118 sp:MALQ_ECOLI Escherichia coli K12 malQ 28.3 55.4 738 4-alpha-glucanotransferase 2520 6020 2432508 2434370 1863 gp:AB005752_1 Lactobacillus brevis plasmid 29.5 64.4 604 ABC transporter, Hop-Resistance horA protein 2521 6021 2433868 2433614 255 GSP:Y74827 Neisseria gonorrhoeae 44.0 51.0 68 Neisserial polypeptides predicted to be useful antigens for vaccines and diagnostics 2522 6022 2434207 2433875 333 GSP:Y74829 Neisseria meningitidis 47.0 53.0 107 polypeptides predicted to be useful antigens for vaccines and diagnostics 2523 6023 2434619 2434440 180 2524 6024 2434776 2434573 204 2525 6025 2436838 2434805 2034 sp:DCP_SALTY Salmonella typhimurium dcp 40.3 68.3 690 peptidyl-dipeptidase 2526 6026 2436871 2438049 1179 gp:AF064523_1 Anisopteromalus calandrae 24.1 45.7 453 carboxylesterase 2527 6027 2438113 2439906 1794 pir:G70983 Mycobacterium tuberculosis 65.2 84.9 594 glycosyl hydrolase or trehalose H37Rv Rv0126 synthase 2528 6028 2439906 2440994 1089 pir:H70983 Mycobacterium tuberculosis 32.1 58.8 449 hypothetical protein H37Rv Rv0127 2529 6029 2441589 2441005 585 pir.T07979 Chlamydomonas reinhardtii ipi1 31.8 57.7 189 isopentenyl-diphosphate Delta- isomerase 2530 6030 2441669 2441890 222 2531 6031 2442355 2442792 438 2532 6032 2443356 2441602 1755 2533 6033 2444015 2443356 660 2534 6034 2444551 2444033 519 2535 6035 2444735 2445709 975 gp:CORCSLYS_1 Corynebacterium glutamicum 99.4 100.0 325 beta C-S lyase (degradation of ATCC 13032 aecD aminoethylcysteine) 2536 6036 2445716 2446993 1278 sp:BRNQ_CORGL Corynebacterium glutamicum 99.8 100.0 426 branched-chain amino acid transport ATCC 13032 brnQ system carrier protein (isoleucine uptake) 2537 6037 2447021 2447998 978 sp:LUXA_VIBHA Vibrio harveyi luxA 21.6 49.0 343 alkanal monooxygenase alpha chain 2538 6038 2450844 2450323 522 2539 6039 2451785 2450859 927 gp:AF155772_2 SinoRhizobium meliloti mdcF 25.9 60.5 324 malonate transporter 2540 6040 2454637 2451794 2844 sp:GLCD_ECOLI Escherichia coli K12 glcD 27.7 55.1 483 glycolate oxidase subunit 2541 6041 2454725 2455435 711 sp:YDFH_ECOLI Escherichia coli K12 ydfH 25.6 65.0 203 transcriptional regulator 2542 6042 2455733 2455452 282 2543 6043 2457066 2455720 1347 sp:YGIK_SALTY Salmonella typhimurium ygiK 22.5 57.6 467 hypothetical protein 2544 6044 2457759 2457337 423 2545 6045 2457863 2459371 1509 sp:HBPA_HAEIN Haemophilus influenzae Rd 27.5 55.5 546 heme-binding protein A precursor HI0853 hbpA (hemin-binding lipoprotein) 2546 6046 2459371 2460336 966 sp:APPB_BACSU Bacillus subtilis 168 appB 40.0 73.3 315 oligopeptide ABC transporter (permease) 2547 6047 2460340 2461167 828 sp:DPPC_ECOLI Escherichia coli K12 dppC 43.2 74.5 271 dipeptide transport system permease protein 2548 6048 2461163 2462599 1437 prf:2306258MR Escherichia coli K12 oppD 37.4 66.4 372 oligopeptide transport ATP-binding protein 2549 6049 2462049 2461543 507 PIR:G72536 Aeropyrum pernix K1 APE1580 35.0 44.0 106 hypothetical protein 2550 6050 2463150 2462602 549 pir:D70367 Aquifex aeolicus VF5 aq_768 29.3 58.0 157 hypothetical protein 2551 6051 2463241 2464143 903 prf:2514301A Rhizobium etli rbsK 41.0 65.0 300 ribose kinase 2552 6052 2464344 2465768 1425 gp:SCM2_16 Streptomyces coelicolor A3(2) 39.9 64.6 466 hypothetical membrane protein SCM2.16c 2553 6053 2465767 2465465 303 2554 6054 2467009 2466038 972 sp:NTCI_HUMAN Homo sapiens 31.3 61.6 284 sodium-dependent transporter or odium Bile acid symporter family 2555 6055 2467077 2467922 846 gp:AF195243_1 Chlamydomonas reinhardtii 28.5 51.2 295 apospory-associated protein C 2556 6056 2470313 2470678 366 2557 6057 2472250 2472819 570 sp:THIX_CORGL Corynebacterium glutamicum 100.0 100.0 133 thiamine biosynthesis protein X ATCC 13032 thiX 2558 6058 2473480 2472893 588 sp:VG66_BPMD Mycobacteriophage D29 66 42.6 65.5 197 hypothetical protein 2559 6059 2473653 2475542 1890 sp:BETP_CORGL Corynebacterium glutamicum 39.8 71.7 601 glycine betaine transporter ATCC 13032 betP 2560 6060 2476497 2477492 996 2561 6061 2477644 2479251 1608 2562 6062 2479379 2479762 384 2563 6063 2481208 2479898 1311 prf:2320266C Rhodobacter capsulatus dctM 34.6 71.9 448 large integral C4-dicarboxylate membrane transport protein 2564 6064 2481692 2481213 480 gp:AF186091_1 Klebsiella pneumoniae dctQ 33.9 73.7 118 small integral C4-dicarboxylate membrane transport protein 2565 6065 2482480 2481734 747 sp:DCTP_RHOCA Rhodobacter capsulatus B10 28.2 59.0 227 C4-dicarboxylate-binding dctP periplasmic protein precursor 2566 6066 2483845 2484087 243 PRF:1806416A Lycopersicon esculentum 63.0 73.0 46 extensin I (tomato) 2567 6067 2484392 2482548 1845 sp:LEPA_BACSU Bacillus subtilis 168 lepA 58.7 83.6 603 GTP-binding protein 2568 6068 2484661 2485269 609 pir:H70683 Mycobacterium tuberculosis 41.6 69.7 185 hypothetical protein H37Rv Rv2405 2569 6069 2485473 2485733 261 sp:RS20_ECOLI Escherichia coli K12 rps T 48.2 72.9 85 30S ribosomal protein S20 2570 6070 2486469 2485801 669 sp:RHTC_ECOLI Escherichia coli K12 rhtC 30.0 67.1 210 thrreonine efflux protein 2571 6071 2486881 2486477 405 gp:SC6D7_25 Streptomyces coelicolor A3(2) 61.2 80.6 129 ankyrin-like protein SC6D7.25. 2572 6072 2487884 2486910 975 pir:H70684 Mycobacterium tuberculosis 46.0 74.1 313 hypothetical protein H37Rv Rv2413c 2573 6073 2489450 2487912 1539 sp:CME3_BACSU Bacillus subtilis 168 comEC 21.4 49.7 527 late competence operon required for DNA binding and uptake 2574 6074 2490154 2489573 582 sp:CME1_BACSU Bacillus subtilis 168 comEA 30.8 63.6 195 late competence operon required for DNA binding and uptake 2575 6075 2490911 2491732 822 2576 6076 2491111 2490290 822 gp:SCC123_7 Streptomyces coelicolor A3(2) 34.8 66.3 273 hypothetical protein SCC123.07c. 2577 6077 2491858 2491151 708 pir:F70685 Mycobacterium tuberculosis 46.8 66.4 235 phosphoglycerate mutase H37Rv Rv2419c 2578 6078 2492343 2491873 471 pir:G70685 Mycobacterium tuberculosis 55.6 86.3 117 hypothetical protein H37Rv Rv2420c 2579 6079 2493178 2492501 678 gp:SCC123_17 Streptomyces coelicolor A3(2) 68.0 85.3 197 hypothetical protein SCC123.17c. 2580 6080 2494237 2493215 1023 2581 6081 2495634 2494339 1296 sp:PROA_CORGL Corynebacterium glutamicum 99.1 99.8 432 gamma-glutamyl phosphate ATCC 17965 proA reductase or glutamate-5- semialdehyde dehydrogenase 2582 6082 2496607 2495696 912 sp:YPRA_CORGL Corynebacterium glutamicum 99.3 100.0 304 D-isomer specific 2-hydroxyacid ATCC 17965 unkdh dehydrogenase 2583 6083 2496803 2497513 711 2584 6084 2499511 2498009 1503 gp:D87915_1 Streptomyces coelicolor A3(2) 58.9 78.2 487 GTP-binding protein obg 2585 6085 2499783 2501669 1887 sp:PBUX_BACSU Bacillus subtilis 168 pbuX 39.1 77.3 422 xanthine permease 2586 6086 2502577 2501735 843 pir:I40838 Corynebacterium sp. ATCC 61.2 81.9 276 2,5-diketo-D-gluconic acid reductase 31090 2587 6087 2502735 2503355 621 2588 6088 2503870 2504265 396 2589 6089 2504247 2503984 264 sp:RL27_STRGR Streptomyces griseus IFO13189 80.3 92.6 81 50S ribosomal protein L27 rpmA 2590 6090 2504602 2504300 303 prf:2304263A Streptomyces griseus IFO13189 56.4 82.2 101 50S ribosomal protein L21 obg 2591 6091 2507098 2504831 2268 sp.RNE_ECOLI Escherichia coli K12 rne 30.1 56.6 886 ribonuclease E 2592 6092 2507115 2507663 549 2593 6093 2507138 2507710 573 2594 6094 2508094 2508840 747 2595 6095 2508922 2509530 609 gp:SCF76_8 Streptomyces coelicolor A3(2) 61.0 82.6 195 hypothetical protein SCF76.08c 2596 6096 2510830 2509523 1308 pir:S43613 Corynebacterium glutamicum 99.1 100.0 436 transposase (insertion sequence ATCC 31831 IS31831) 2597 6097 2511046 2511423 378 gp:SCF76_8 Streptomyces coelicolor A3(2) 51.3 76.9 117 hypothetical protein SCF76.08c 2598 6098 2511427 2511876 450 gp:SCF76_9 Streptomyces coelicolor A3(2) 37.8 67.8 143 hypothetical protein SCF76.09 2599 6099 2512356 2511949 408 gp:AF069544_1 Mycobacterium smegmatis ndk 70.9 89.6 134 nucleoside diphosphate kinase 2600 6100 2512768 2512409 360 2601 6101 2512803 2513144 342 gp:AE002024_10 Deinococcus radiodurans R1 34.8 67.4 92 hypothetical protein DR1844 2602 6102 2513618 2513154 465 pir:H70515 Mycobacterium tuberculosis 36.6 64.3 112 hypothetical protein H37Rv Rv1883c 2603 6103 2514114 2513692 423 pir:E70863 Mycobacterium tuberculosis 33.9 68.6 118 hypothetical protein H37Rv Rv2446c 2604 6104 2515487 2514114 1374 prf:2410252B Streptomyces coelicolor A3(2) 55.4 79.6 451 folyl-polyglutamate synthetase folC 2605 6105 2515662 2516273 612 2606 6106 2516243 2516956 714 2607 6107 2517089 2517751 663 2608 6108 2518336 2515637 2700 sp:SYV_BACSU Bacillus subtilis 168 balS 45.5 72.1 915 valyl-tRNA synthetase 2609 6109 2519972 2518398 1575 pir:A38447 Bacillus subtilis 168 oppA 24.2 58.5 521 oligopeptide ABC transport system substrate-binding protein 2610 6110 2520209 2521660 1452 sp:DNAK_BACSU Bacillus subtilis 168 dnaK 26.2 54.9 508 heat shock protein dnaK 2611 6111 2522251 2521667 585 gp:ECU89166_1 Eikenella corrodens ATCC 42.9 71.2 170 lysine decarboxylase 23824 2612 6112 2523248 2522265 984 sp:MDH_THEFL Thermus aquaticus ATCC 33923 56.4 76.5 319 malate dehydrogenase mdh 2613 6113 2523561 2524337 777 gp:SC4A10_33 Streptomyces coelicolor A3(2) 24.6 56.5 207 transcriptional regulator SC4A10.33 2614 6114 2524915 2524340 576 gp:AF065442_1 Vibrio cholerae aphA 26.0 51.4 208 hypothetical protein 2615 6115 2525099 2526226 1128 prf:2513416F Acinetobacter sp. vanA 39.5 68.6 357 vanillate demethylase(oxygenase) 2616 6116 2526233 2527207 975 gp:FSU12290_2 Sphingomonas flava ATCC 32.8 59.2 338 pentachlorophenol 4- 39723 pcpD monooxygenase reductase 2617 6117 2527135 2528559 1425 prf:2513416G Acinetobacter sp. vanK 40.8 76.8 444 transport protein 2618 6118 2529480 2528551 930 gp:KPU95087_7 Klebsiella pneumoniae mdcF 28.0 58.4 286 malonate transporter 2619 6119 2530761 2529484 1278 prf:2303274A Bacillus subtilis clpX 59.8 85.8 430 class-III heat-shock protein or ATP- dependent protease 2620 6120 2530891 2531976 1086 gp:SCF55_28 Streptomyces coelicolor A3(2) 45.6 73.0 366 hypothetical protein SCF55.28c 2621 6121 2532601 2531969 633 gp:AF109386_2 Streptomyces sp. 2065 pcaJ 63.3 85.7 210 succinyl CoA:3-oxoadipate CoA transferase beta subunit 2622 6122 2533353 2532604 750 gp:AF109386_1 Streptomyces sp. 2065 pcal 60.2 84.5 251 succinyl CoA:3-oxoadipate CoA transferase alpha subunit 2623 6123 2533391 2534182 792 prf:2408324F Rhodococcus opacus 1CP pcaR 58.2 82.5 251 protocatechuate catabolic protein 2624 6124 2534201 2535424 1224 prf:2411305D Ralstonia eutropha bktB 44.8 71.9 406 beta-ketothiolase 2625 6125 2535168 2534257 912 2626 6126 2535430 2536182 753 prf:2408324E Rhodococcus opacus pcaL 50.8 76.6 256 3-oxoadipate enol-lactone hydrolase and 4-carboxymuconolactone decarboxylase 2627 6127 2536196 2538256 2061 gp:SCM1_10 Streptomyces coelicolor A3(2) 23.6 43.0 825 transcriptional regulator SCM1.10 2628 6128 2538613 2538248 366 prf:2408324E Rhodococcus opacus pcaL 78.3 89.6 115 3-oxoadipate enol-lactorie hydrolase and 4-carboxymuconolactone decarboxylase 2629 6129 2539553 2540230 678 2630 6130 2539731 2538616 1116 prf:2408324D Rhodococcus opacus pcaB 39.8 63.4 437 3-carboxy-cis, cis-muconate cycloisomerase 2631 6131 2540320 2539709 612 prf:2408324C Rhodococcus opacus pcaG 49.5 70.6 214 protocatechuate dioxygenase alpha subunit 2632 6132 2541024 2540335 690 prf:2408324B Rhodococcus opacus pcaH 74.7 91.2 217 protocatechuate dioxygenase beta subunit 2633 6133 2542350 2541187 1164 pir:G70506 Mycobacterium tuberculosis 26.4 48.7 273 hypothetical protein H37Rv Rv0336 2634 6134 2542802 2542512 291 prf:2515333B Mycobacterium tuberculosis 54.4 81.5 92 muconolactone isomerase catC 2635 6135 2543043 2543813 771 2636 6136 2543936 2542818 1119 sp:CATB_RHOOP Rhodococcus opacus 1CP catB 60.8 84.7 372 muconate cycloisomerase 2637 6137 2544262 2544867 606 2638 6138 2544876 2544022 855 prf:2503218A Rhodococcus rhodochrous catA 72.3 88.4 285 catechol 1,2-dioxygenase 2639 6139 2545068 2544928 141 2640 6140 2545315 2546784 1470 gp:AF134348_1 Pseudomonas putida plasmid 62.2 85.6 437 toluate 1,2 dioxygenase subunit pDK1 xylX 2641 6141 2546827 2547318 492 gp:AF134348_2 Pseudomonas putida plasmid 60.3 83.2 161 toluate 1,2 dioxygenase subunit pDK1 xylY 2642 6142 2547333 2548868 1536 gp:AF134348_3 Pseudomonas putida plasmid 51.5 81.0 342 toluate 1,2 dioxygenase subunit pDK1 xylZ 2643 6143 2548868 2549695 828 gp:AF134348_4 Pseudomonas putida plasmid 30.7 61.4 277 1,2-dihydroxycyclohexa-3,5-diene pDK1 xylL carboxylate dehydrogenase 2644 6144 2549771 2552455 2685 gp:REU95170_1 Rhodococcus erythropolis thcG 23.3 48.6 979 regulator of LuxR family with ATP- binding site 2645 6145 2552563 2553942 1380 sp:PCAK_ACICA Acinetobacter calcoaceticus 31.3 64.4 435 transmembrane transport protein or pcaK 4-hydroxybenzoate transporter 2646 6146 2554026 2555267 1242 sp:BENE_ACICA Acinetobacter calcoaceticus 29.9 66.2 388 benzoate membrane transport benE protein 2647 6147 2555940 2555317 624 gp:AF071885_2 Streptomyces coelicolor M145 69.5 88.3 197 ATP-dependent Clp protease clpP2 proteolytic subunit 2 2648 6148 2556580 2555978 603 gp:AF071885_1 Streptomyces coelicolor M145 62.1 85.9 198 ATP-dependent Clp protease clpP1 proteolytic subunit 1 2649 6149 2556599 2556748 150 gp:SIS243537_4 Sulfolobus islandicus ORF154 42.9 71.4 42 hypothetical protein 2650 6150 2558106 2556760 1347 sp:TIG_BACSU Bacillus subtilis 168 tig 32.1 66.4 417 trigger factor (prolyl isomerase) (chaperone protein) 2651 6151 2558609 2559103 495 gp:SCD25_17 Streptomyces coelicolor A3(2) 32.5 63.1 160 hypothetical protein SCD25.17 2652 6152 2559157 2560131 975 sp:PBP4_NOCLA Nocardia lactamdurans LC411 25.3 50.9 336 penicillin-binding protein pbp 2653 6153 2560131 2560586 456 prf:2301342A Mus musculus Moa1 27.8 58.3 115 hypothetical protein 2654 6154 2561115 2561363 249 2655 6155 2561920 2561483 438 prf:2513302C Corynebacterium striatum ORF1 54.2 73.2 142 transposase 2656 6156 2562093 2562242 150 2657 6157 2562115 2561990 126 prf:2513302C Corynebacterium striatum ORF1 57.1 82.9 35 hypothetical protein 2658 6158 2562341 2562078 264 prf:2513302C Corynebacterium striatum ORF1 50.7 78.7 75 transposase 2659 6159 2562776 2562387 390 2660 6160 2562963 2563847 885 2661 6161 2564402 2563932 471 sp:LACB_STAAU Staphylococcus aureus NCTC 40.0 71.4 140 galactose-6-phosphate isomerase 8325-4 lacB 2662 6162 2565245 2564550 696 sp:YAMY_BACAD Bacillus acidopullulyticus ORF2 26.2 58.1 248 hypothetical protein 2663 6163 2566231 2565623 609 pir:A70866 Mycobacterium tuberculosis 56.8 80.9 199 hypothetical protein H37Rv Rv2466c 2664 6164 2566345 2568945 2601 sp:AMPN_STRLI Streptomyces lividans pepN 47.5 70.5 890 aminopeptidase N 2665 6165 2569211 2570293 1083 pir:B70206 Borrelia burgdorferi BB0852 25.1 58.1 358 hypothetical protein 2666 6166 2571460 2570309 1152 2667 6167 2571510 2572175 666 2668 6168 2572193 2572348 156 2669 6169 2572677 2572351 327 gp:AF139916_3 Brevibacterium linens ATCC 61.5 81.7 104 phytoene desaturase 9175 crtl 2670 6170 2572977 2572807 171 2671 6171 2573770 2573393 378 2672 6172 2573864 2572659 1206 sp:CRTJ_MYXXA Myxococcus xanthus DK1050 31.2 63.8 381 phytoene dehydrogenase carA2 2673 6173 2574718 2573843 876 sp:CRTB_STRGR Streptomyces griseus JA3933 31.4 58.6 290 phytoene synthase crtB 2674 6174 2575898 2574780 1119 gp:LMAJ9627_3 Listeria monocytogenes IltB 25.8 47.7 392 multidrug resistance transporter 2675 6175 2577213 2575981 1233 2676 6176 2578872 2577232 1641 gp:SYOATPBP_2 Synechococcus elongatus 41.3 71.6 538 ABC transporter ATP-binding protein 2677 6177 2579760 2578879 882 sp:DPPC_BACFI Bacillus firmus OF4 dppC 38.8 73.8 286 dipeptide transport system permease protein 2678 6178 2580707 2579769 939 pir:S47696 Escherichia coli K12 nikB 33.2 62.0 316 nickel transport system permease protein 2679 6179 2582417 2580711 1707 2680 6180 2582564 2584504 1941 2681 6181 2584613 2585926 1314 sp:ARGD_CORGL Corynebacterium glutamicum 31.4 63.5 411 acetylornithine aminotransferase ATCC 13032 argD 2682 6182 2586180 2587763 1584 pir:A70539 Mycobacterium tuberculosis 25.1 47.9 482 hypothetical protein H37Rv Rv1128c 2683 6183 2587976 2588722 747 sp:YA26_MYCTU Mycobacterium tuberculosis 49.1 79.4 218 hypothetical membrane protein H37Rv Rv0364 2684 6184 2589432 2588725 708 sp:PHBB_CHRVI Chromatium vinosum D phbB 28.1 60.0 235 acetoacetyl CoA reductase 2685 6185 2589565 2590302 738 pir:A40046 Streptomyces coelicolor actll 26.7 55.0 240 transcriptional regulator, TetR family 2686 6186 2590697 2591137 441 GSP:Y74375 Neisseria meningitidis 38.0 47.0 94 polypeptides predicted to be useful antigens for vaccines and diagnostics 2687 6187 2592365 2591574 792 gp:AF106002_1 Pseudomonas putida GM73 31.1 65.1 238 ABC transporter ATP-binding protein ttg2A 2688 6188 2592402 2592794 393 gp:MLCB1610_9 Mycobacterium leprae 53.2 77.0 126 globin MLCB1610.14c 2689 6189 2592838 2593965 1128 sp:CHRA_PSEAE Pseudomonas aeruginosa 27.3 60.4 396 chromate transport protein Plasmid pUM505 chrA 2690 6190 2594594 2593968 627 pir:A70867 Mycobacterium tuberculosis 37.8 68.9 196 hypothetical protein H37Rv Rv2474c 2691 6191 2595061 2594597 465 gp:SC6D10_19 Streptomyces coelicolor A3(2) 36.2 61.4 127 hypothetical protein SC6D10.19c 2692 6192 2595808 2595188 621 2693 6193 2595983 2595822 162 pir:B72589 Aeropyrum pernix K1 APE1182 36.4 60.0 55 hypothetical protein 2694 6194 2597715 2596048 1668 sp:YJJK_ECOLI Escherichia coli K12 yjjK 52.8 79.6 563 ABC transporter ATP-binding protein 2695 6195 2598483 2597869 615 pir:E70867 Mycobacterium tuberculosis 31.4 62.2 172 hypothetical protein H37Rv Rv2478c 2696 6196 2600764 2598662 2103 sp:Y05L_MYCLE Mycobacterium leprae 0659 28.0 56.7 700 hypothetical membrane protein 2697 6197 2601461 2602879 1419 pir:C69676 Bacillus subtilis phoB 28.0 52.6 536 alkaline phosphatase 2698 6198 2604573 2605502 930 2699 6199 2604583 2603945 639 2700 6200 2605520 2604609 912 sp:MSMG_STRMU Streptococcus mutans 39.1 76.3 279 multiple sugar-binding transport INGBRITTmsmG system permease protein 2701 6201 2606369 2605527 843 sp:MSMF_STRMU Streptococcus mutans 27.4 67.5 292 multiple sugar-binding transport INGBRITTmsmF system permease protein 2702 6202 2606444 2608117 1674 2703 6203 2607889 2606561 1329 prf:2206392C Thermoanaerobacterium 28.8 63.2 462 maltose-binding protein thermosul amyE 2704 6204 2609426 2608185 1242 2705 6205 2610639 2609512 1128 prf:2308356A Streptomyces reticuli msiK 59.1 79.8 386 ABC transporter ATP-binding protein (ABC-type sugar transport protein) or cellobiose/maltose transport protein 2706 6206 2611523 2612272 750 2707 6207 2611531 2610848 684 prf:2317468A Schizosaccharomyces pombe 37.7 72.7 154 dolichol phosphate mannose dpm1 synthase 2708 6208 2612462 2613151 690 2709 6209 2613712 2614500 789 prf:2516398E Rhodococcus rhodochrous 67.2 89.4 207 aldehyde dehydrogenase plasmid pRTL1 orf5 2710 6210 2614649 2615410 762 prf:2513418A Synechococcus sp. PCC7942 48.6 73.8 183 circadian phase modifier cpmA 2711 6211 2615451 2615795 345 2712 6212 2617120 2615939 1182 pir:A72312 Thermotoga maritima MSB8 35.0 64.6 412 hypothetical membrane protein TM0964 2713 6213 2617246 2617995 750 sp:GIP_ECOLI Escherichia coli K12 gip 41.2 69.4 255 glyoxylate-induced protein 2714 6214 2618072 2618869 798 pir:E70761 Mycobacterium tuberculosis 40.0 57.0 258 ketoacyl reductase H37Rv Rv1544 2715 6215 2618882 2619538 657 sp:ORN_ECOLI Escherichia coli K12 orn 48.0 78.8 179 oligoribonuclease 2716 6216 2620728 2619541 1188 prf:2409378A Salmonella enterica iroD 26.0 50.9 454 ferric enterochelin esterase 2717 6217 2622181 2620973 1209 pir:C70870 Mycobacterium tuberculosis 48.5 71.9 398 lipoprotein H37Rv Rv2518c lppS 2718 6218 2622961 2623605 645 2719 6219 2623770 2623621 150 2720 6220 2623803 2624048 246 2721 6221 2625358 2624051 1308 gp:SCU53587_1 Corynebacterium glutamicum 99.5 99.8 436 transposase (IS1207) ATCC 21086 2722 6222 2625600 2625806 207 2723 6223 2626447 2625809 639 2724 6224 2627924 2628376 453 gp:AF085239_1 Salmonella typhimurium KP1001 32.8 63.4 131 transcriptional regulator cytR 2725 6225 2628121 2626493 1629 sp:GLSK_RAT Rattus norvegicus SPRAGUE- 35.2 69.3 358 glutaminase DAWLEY KIDNEY 2726 6226 2628376 2628852 477 pir:A36940 Bacillus subtilis 168 degA 42.3 72.2 97 sporulation-specific degradation regulator protein 2727 6227 2628878 2628324 555 2728 6228 2628926 2630479 1554 sp:UXAC_ECOLI Escherichia coli K12 uxaC 29.0 60.9 335 uronate isomerase 2729 6229 2630636 2631136 501 2730 6230 2631270 2632466 1197 prf:1814452C Zea diploperennis perennial 32.0 45.0 291 hypothetical protein teosinte 2731 6231 2632543 2633100 558 prf:2324444A Mycobacterium avium pncA 48.1 74.6 185 pyrazinamidase/nicotinamidase 2732 6232 2633418 2633146 273 pir:E70870 Mycobacterium tuberculosis 42.7 80.0 75 hypothetical protein H37Rv Rv2520c 2733 6233 2633600 2634064 465 sp:BCP_ECOLI Escherichia coli K12 bcp 46.8 73.8 141 bacterioferritin comigratory protein 2734 6234 2634116 2634751 636 gp:SCI11_1 Streptomyces coelicolor A3(2) 32.5 61.4 114 bacterial regulatory protein, tetR SCI11.01c family 2735 6235 2635151 2634747 405 gp:BAY15081_1 Corynebacterium 56.6 75.9 145 phosphopantethiene protein ammoniagenes ATCC 6871 ppt1 transferase 2736 6236 2636589 2635165 1425 gp:AF237667_1 Corynebacterium glutamicum 52.4 85.6 473 lincomycin resistance protein lmrB 2737 6237 2636845 2637168 324 pir:S76537 Synechocystis sp. PCC6803 30.1 54.0 113 hypothetical membrane protein 2738 6238 2637653 2637240 414 2739 6239 2647627 2638649 8979 pir:S2047 Corynebacterium 62.3 83.6 3029 fatty-acid synthase ammoniagenes fas 2740 6240 2649416 2648235 1182 gp:SC4A7_14 Streptomyces coelicolor A3(2) 25.3 55.2 404 hypothetical protein SC4A7.14 2741 6241 2649550 2650164 615 pir:D70716 Mycobacterium tuberculosis 40.4 60.9 230 peptidase H37Rv Rv0950c 2742 6242 2650441 2650902 462 sp:Y077_MYCT Mycobacterium tuberculosis 40.2 67.9 112 hypothetical membrane protein H37Rv Rv1343c 2743 6243 2650986 2651339 354 sp:Y076_MYCLE Mycobacterium leprae 37.2 69.0 113 hypothetical membrane protein B1549_F2_59 2744 6244 2652037 2651420 618 sp:Y03Q_MYCTU Mycobacterium tuberculosis 55.0 76.7 202 hypothetical protein H37Rv Rv1341 2745 6245 2652801 2652067 735 sp:RNPH_PSEAE Pseudomonas aeruginosa 60.2 81.4 236 ribonuclease PH ATCC 15692 rph 2746 6246 2653254 2653009 246 2747 6247 2654018 2653326 693 2748 6248 2654660 2654079 582 2749 6249 2656236 2654875 1362 sp:Y029_MYCTU Mycobacterium tuberculosis 29.0 58.2 428 hypothetical membrane protein H37Rv SC8A6.09c 2750 6250 2656452 2656985 534 gp:AF121000_8 Corynebacterium glutamicum 92.1 97.2 175 transposase (IS1628) 22243 R-plasmid pAG1 tnpB 2751 6251 2657633 2656974 660 2752 6252 2658500 2657736 765 sp:Y03O_MYCLE Mycobacterium leprae ats 46.0 74.4 250 arylsulfatase 2753 6253 2659457 2658606 852 prf:2516259A Corynebacterium glutamicum 99.3 99.3 284 D-glutamate racemase ATCC 13869 murI 2754 6254 2659496 2660131 636 2755 6255 2660638 2660147 492 gp:SCE22_22 Streptomyces coelicolor A3(2) 44.2 70.8 147 bacterial regulatory protein, marR SCE22.22 family 2756 6256 2661417 2660671 747 sp:Y03M_MYCTU Mycobacterium tuberculosis 38.2 69.3 225 hypothetical membrane protein H37Rv Rv1337 2757 6257 2661565 2662455 891 2758 6258 2662376 2661417 960 pir:A47039 Flavobacterium sp. nylC 30.2 58.3 321 endo-type 6-aminohexanoate oligomer hydrolase 2759 6259 2662867 2662331 537 sp:Y03H_MYCTU Mycobacterium tuberculosis 35.0 58.5 200 hypothetical protein H37Rv Rv1332 2760 6260 2663182 2662883 300 sp:Y03G_MYCTU Mycobacterium tuberculosis 57.1 77.1 105 hypothetical protein H37Rv Rv1331 2761 6261 2663437 2664060 624 2762 6262 2664060 2665397 1338 sp:Y03F_MYCTU Mycobacterium tuberculosis 61.2 80.8 428 hypothetical protein H37Rv Rv1330c 2763 6263 2665687 2665992 306 2764 6264 2666115 2667854 1740 prf:1816252A Escherichia coli dinG 25.2 53.3 647 ATP-dependent helicase 2765 6265 2668760 2667870 891 sp:Y0A8_MYCTU Mycobacterium tuberculosis 29.7 60.1 313 hypothetical membrane protein H37Rv Rv2560 2766 6266 2669561 2668839 723 pir:T34684 Streptomyces coelicolor A3(2) 39.0 52.0 222 hypothetical protein SC1B5.06c 2767 6267 2670573 2669557 1017 sp:SERB_ECOLI Escherichia coli K12 serB 38.7 61.0 310 phosphoserine phosphatase 2768 6268 2671126 2672721 1596 2769 6269 2672805 2671063 1743 pir:D45335 Mycobacterium tuberculosis 46.8 74.4 575 cytochrome c oxidase chain I H37Rv Rv3043c 2770 6270 2672950 2673255 306 2771 6271 2674339 2673338 1002 gp:AF112536_1 Corynebacterium glutamicum 99.7 99.7 334 ribonucleotide reductase beta-chain ATCC 13032 nrdF 2772 6272 2674804 2675289 486 sp:FTNA_ECOLI Escherichia coli K12 ftnA 31.5 64.2 159 ferritin 2773 6273 2675491 2676240 750 gp:SCA32WHIH_4 Streptomyces coelicolor A3(2) 32.8 60.2 256 sporulation transcription factor whiH 2774 6274 2676902 2676243 660 pir:I40339 Corynebacterium glutamicum 27.6 60.4 225 iron dependent repressor or ATCC 13869 dtxR diptheria toxin repressor 2775 6275 2676940 2677377 438 sp:TIR2_YEAST Saccharomyces cerevisiae 24.2 62.1 124 cold shock protein TIR2 precursor YPH148 YOR010C TIR2 2776 6276 2677193 2676918 276 pir:C69281 Archaeoglobus fulgidus AF0251 50.0 86.0 50 hypothetical membrane protein 2777 6277 2679598 2677478 2121 gp:AF112535_3 Corynebacterium glutamicum 99.9 100.0 707 ribonucleotide reductase alpha- ATCC 13032 nrdE chain 2778 6278 2680470 2680784 315 2779 6279 2681363 2681223 141 SP:RL36_RICPR Rickettsia prowazekii 58.0 79.0 41 50S ribosomal protein L36 2780 6280 2681546 2682376 831 sp:NADE_BACSU Bacillus subtilis 168 nadE 55.6 78.1 279 NH3-dependent NAD(+) synthetase 2781 6281 2681556 2681464 93 2782 6282 2683119 2683616 498 2783 6283 2683125 2682379 747 pir:S76790 Synechocystis sp. PCC6803 30.7 56.4 257 hypothetical protein slr1563 2784 6284 2683418 2683131 288 pir:G70922 Mycobacterium tuberculosis 41.7 68.8 96 hypothetical protein H37Rv Rv3129 2785 6285 2684646 2683627 1020 sp:ADH2_BACST Bacillus stearothermophilus 26.1 52.8 337 alcohol dehydrogenase DSM 2334 adh 2786 6286 2684919 2686289 1371 sp:MMGE_BACSU Bacillus subtilis 168 mmgE 27.0 56.0 459 Bacillus subtilis mmg (for mother cell metabolic genes) 2787 6287 2686315 2687148 834 pir:T05174 Arabidopsis thaliana T6K22.50 33.8 66.2 284 hypothetical protein 2788 6288 2688240 2687449 792 2789 6289 2690050 2688389 1662 sp:PGMU_ECOLI Escherichia coli K12 pgm 61.7 80.6 556 phosphoglucomutase 2790 6290 2690150 2690437 288 pir:F70650 Mycobacterium tuberculosis 41.7 64.3 84 hypothetical membrane protein H37Rv Rv3069 2791 6291 2690437 2690760 324 pir:D71843 Helicobacter pylon J99 jhp1146 25.4 61.5 122 hypothetical membrane protein 2792 6292 2690773 2691564 792 sp:YCSI_BACSU Bacillus subtilis 168 ycsl 51.2 79.1 254 hypothetical protein 2793 6293 2691689 2693053 1365 gp:AF126281_1 Rhodococcus erythropolis 24.2 48.6 496 transposase (IS1676) 2794 6294 2693299 2694918 1620 sp:CSP1_CORGL Corynebacterium glutamicum 24.8 49.6 355 major secreted protein PS1 protein (Brevibacterium flavum) ATCC precursor 17965 csp1 2795 6295 2694926 2695279 354 2796 6296 2695554 2695718 165 2797 6297 2695766 2695320 447 2798 6298 2695812 2697212 1401 gp:AF126281_1 Rhodococcus erythropolis 24.6 46.6 500 transposase (IS1676) 2799 6299 2698150 2697383 768 2800 6300 2699531 2698194 1338 sp:GLTT_BACCA Bacillus subtilis 168 30.8 66.2 438 proton/sodium-glutamate symport protein 2801 6301 2700920 2701612 693 2802 6302 2702466 2699926 2541 gp:SCE25_30 Streptomyces coelicolor A3(2) 33.0 69.0 873 ABC transporter SCE25.30 2803 6303 2702466 2703356 891 2804 6304 2703194 2702487 708 gp:SAU18641_2 Staphylococcus aureus 45.4 79.8 218 ABC transporter ATP-binding protein 2805 6305 2704314 2704586 273 PIR:F81516 Chlamydophila pneumoniae 60.0 67.0 84 hypothetical protein AR39 CP0987 2806 6306 2704835 2704975 141 PIR:F81737 Chlamydia muridarum Nigg 71.0 75.0 42 hypothetical protein TC0129 2807 6307 2709878 2710555 678 2808 6308 2710637 2711308 672 prf:2509388L Streptomyces collinus Tu 1892 28.1 54.1 196 oxidoreductase or dehydrogenase ansG 2809 6309 2711850 2712374 525 sp:Y089_MYCTU Mycobacterium tuberculosis 25.9 51.2 205 methyltransferase H37Rv Rv0089 2810 6310 2713181 2713453 273 GSP:Y35814 Chlamydia pneumoniae 61.0 66.0 84 hypothetical protein 2811 6311 2713702 2713842 141 PIR:F81737 Chlamydia muridarum Nigg 71.0 75.0 42 hypothetical protein TC0129 2812 6312 2718187 2717993 195 2813 6313 2719689 2718436 1254 sp:MURA_ACICA Acinetobacter calcoaceticus 44.8 75.3 417 UDP-N-acetylglucosamine 1- NCIB 8250 murA carboxyvinyltransferase 2814 6314 2719750 2720319 570 sp:Y02Y_MYCTU Mycobacterium tuberculosis 66.3 84.2 190 hypothetical protein H37Rv Rv1314c 2815 6315 2721227 2720385 843 gp:SC2G5_15 Streptomyces coelicolor A3(2) 45.9 69.0 281 transcriptional regulator SC2G5.15c 2816 6316 2721702 2721295 408 2817 6317 2721934 2722857 924 sp:CYSK_BACSU Bacillus subtilis 168 cysK 57.1 84.6 305 cysteine synthase 2818 6318 2723064 2723609 546 prf:2417357C Azotobacter vinelandii cysE2 61.1 79.7 172 O-acetylserine synthase 2819 6319 2724057 2723770 288 gp:AE002024_10 Deinococcus radiodurans R1 36.1 65.1 83 hypothetical protein DR1844 2820 6320 2725359 2724478 882 sp:SUCD_COXBU Coxiella burnetii Nine Mile Ph I 52.9 79.4 291 succinyl-CoA synthetase alpha sucD chain 2821 6321 2725619 2725843 225 PIR:F72706 Aeropyrum pernix K1 APE1069 42.0 43.0 75 hypothetical protein 2822 6322 2726577 2725384 1194 sp:SUCC_BACSU Bacillus subtilis 168 sucC 39.8 73.0 400 succinyl-CoA synthetase beta chain 2823 6323 2727145 2726786 360 2824 6324 2728133 2727399 735 gp:AF058302_5 Streptomyces roseofulvus frnE 38.5 71.8 213 frenolicin gene E product 2825 6325 2729025 2728207 819 2826 6326 2730916 2729378 1539 sp:CAT1_CLOKL Clostridium kluyveri cat1 cat1 47.9 77.8 501 succinyl-CoA coenzyme A transferase 2827 6327 2731376 2732518 1143 sp:NIR3_AZOBR Azospirillum brasilense ATCC 38.6 68.5 321 transcriptional regulator 29145 ntrC 2828 6328 2732230 2731424 807 2829 6329 2732636 2733367 732 pir:E70810 Mycobacterium tuberculosis 46.5 81.7 213 phosphate transport system H37Rv Rv0821c phoY-2 regulatory protein 2830 6330 2734351 2733455 897 pir:S68595 Pseudomonas aeruginosa pstB 58.8 82.8 255 phosphate-specific transport component 2831 6331 2735184 2734264 921 gp:MTPSTA1_1 Mycobacterium tuberculosis 51.4 82.2 292 phosphate ABC transport system H37Rv Rv0830 pstA1 permease protein 2832 6332 2736215 2735202 1014 pir:A70584 Mycobacterium tuberculosis 50.2 78.5 325 phosphate ABC transport system H37Rv Rv0829 pstC2 permease protein 2833 6333 2737538 2736414 1125 pir:H70583 Mycobacterium tuberculosis 40.0 56.0 369 phosphate-binding protein S-3 H37Rv phoS2 precursor 2834 6334 2738711 2737836 876 gp:SCD84_18 Streptomyces coelicolor A3(2) 34.3 60.0 315 acetyltransferase SCD84.18c 2835 6335 2738771 2739553 783 2836 6336 2740650 2739556 1095 sp:BMRU_BACSU Bacillus subtilis 168 bmrU 24.7 55.2 344 hypothetical protein 2837 6337 2740670 2741356 687 pir:E70809 Mycobacterium tuberculosis 44.9 74.2 225 hypothetical protein H37Rv Rv0813c 2838 6338 2742577 2741636 942 gp:AF193846_1 Solanum tuberosum BCAT2 28.6 56.0 259 branched-chain amino acid aminotransferase 2839 6339 2742685 2743785 1101 gp:AB003158_6 Corynebacterium 58.5 79.0 352 hypothetical protein ammoniagenes ATCC 6872 ORF4 2840 6340 2744010 2744222 213 pir:B70809 Mycobacterium tuberculosis 58.6 81.0 58 hypothetical protein H37Rv Rv0810c 2841 6341 2745954 2744881 1074 gp:AB003158_5 Corynebacterium 81.0 94.2 347 5′-phosphoribosyl-5-aminoimidazole ammoniagenes ATCC 6872 synthetase purM 2842 6342 2747564 2746083 1482 gp:AB003158_4 Corynebacterium 70.3 89.0 482 amidophosphoribosyl transferase ammoniagenes ATCC 6872 purF 2843 6343 2748057 2747683 375 pir:H70536 Mycobacterium tuberculosis 57.3 75.8 124 hypothetical protein H37Rv Rv0807 2844 6344 2748095 2749111 1017 gp:AB003158_2 Corynebacterium 75.9 94.0 315 hypothetical protein ammoniagenes ATCC 6872 ORF2 2845 6345 2749902 2749162 741 gp:AB003158_1 Corynebacterium 67.7 87.1 217 hypothetical membrane protein ammoniagenes ATCC 6872 ORF1 2846 6346 2751918 2752103 186 GP:SSU18930_21 Sulfolobus solfataricus 64.0 71.0 42 hypothetical protein 4 2847 6347 2752312 2750027 2286 gp:AB003162_3 Corynebacterium 77.6 89.5 763 5′-phosphoribosyl-N- ammoniagenes ATCC 6872 formylglycinamidine synthetase purL 2848 6348 2752402 2753121 720 2849 6349 2752995 2752327 669 gp:AB003162_2 Corynebacterium 80.3 93.3 223 5′-phosphoribosyl-N- ammoniagenes ATCC 6872 formylglycinamidine synthetase purQ 2850 6350 2753237 2752995 243 gp:AB003162_1 Corynebacterium 81.0 93.7 79 hypothetical protein ammoniagenes ATCC 6872 purorf 2851 6351 2753298 2753819 522 2852 6352 2753804 2753328 477 prf:2420329A Lactococcus lactis gpo 46.2 77.9 158 gluthatione peroxidase 2853 6353 2753992 2756739 2748 prf:2216389A Aeromonas hydrophila JMP636 28.0 51.5 965 extracellular nuclease nucH 2854 6354 2756851 2757126 276 2855 6355 2757815 2757129 687 pir:C70709 Mycobacterium tuberculosis 37.4 68.7 211 hypothetical protein H37Rv Rv0784 2856 6356 2759200 2757863 1338 sp:DCTA_SALTY Salmonella typhimurium LT2 49.0 81.6 414 C4-dicarboxylate transporter dctA 2857 6357 2761649 2759532 2118 prf:2408266A Pseudomonas sp. WO24 dapb1 41.8 70.6 697 dipeptidyl aminopeptidase 2858 6358 2762452 2761829 624 2859 6359 2762675 2761785 891 gp:AB003161_3 Corynebacterium 70.1 89.1 294 5′-phosphoribosyl-4-N- ammoniagenes ATCC 6872 succinocarboxamide-5-amino purC imidazole synthetase 2860 6360 2764931 2763504 1428 gp:AB003161_2 Corynebacterium 85.3 95.0 477 adenylosuccino lyase ammoniagenes ATCC 6872 purB 2861 6361 2766135 2764978 1158 sp:AAT_SULSO Sulfolobus solfataricus ATCC 28.1 62.3 395 aspartate aminotransferase 49255 2862 6362 2767420 2766158 1263 gp:AB003161_1 Corynebacterium 71.1 86.4 425 5′-phosphoribosylglycinamide ammoniagenes ATCC 6872 synthetase purD 2863 6363 2767580 2767993 414 sp:YHIT_MYCLE Mycobacterium leprae u296a 53.7 80.2 136 histidine triad (HIT) family protein 2864 6364 2768137 2767703 435 2865 6365 2769095 2768343 753 pir:S62195 Methanosarcina barkeri orf3 26.8 56.4 243 hypothetical protein 2866 6366 2770511 2769156 1356 sp:DTPT_LACLA Lactococcus lactis subsp. lactis 30.1 67.6 469 di-/tripeptide transpoter dipT 2867 6367 2770714 2771982 1269 sp:BIOA_CORGL Corynebacterium glutamicum 95.7 98.8 423 adenosylmethionine-8-amino-7- (Brevibacterium flavum) MJ233 oxononanoate aminotransferase or bioA 7,8-diaminopelargonic acid aminotransferase 2868 6368 2771989 2772660 672 sp:BIOD_CORGL Corynebacterium glutamicum 98.7 99.6 224 dethiobiotin synthetase (Brevibacterium flavum) MJ233 bioD 2869 6369 2774098 2772644 1455 gp:AF049873_3 Lactococcus lactis M71plasmid 31.3 70.5 335 two-component system sensor pND306 histidine kinase 2870 6370 2774814 2774110 705 prf:2222216A Thermotoga maritima drrA 42.0 72.7 231 two-component system regulatory protein 2871 6371 2775689 2774937 753 sp:TIPA_STRLI Streptomyces lividans tipA 37.4 69.5 249 transcriptional activator 2872 6372 2776879 2775740 1140 prf:2419350A Arthrobacter sp. DK-38 30.9 53.9 382 metal-activated pyridoxal enzyme or low specificity D-Thr aldolase 2873 6373 2778504 2776768 1737 gp:ECOPOXB8G_1 Escherichia coli K12 poxB 46.3 75.8 574 pyruvate oxidase 2874 6374 2778965 2780446 1482 prf.2212334B Staphylococcus aureus plasmid 33.3 68.9 504 multidrug efflux protein pSK23 qacB 2875 6375 2780439 2780969 531 sp:YCDC_ECOLI Escherichia coli K12 ycdC 30.4 68.5 92 transcriptional regulator 2876 6376 2780996 2782315 1320 pir:D70551 Mycobacterium tuberculosis 45.6 78.4 421 hypothetical membrane protein H37Rv Rv2508c 2877 6377 2784481 2782340 2142 2878 6378 2785615 2784656 960 gp.AF096929_2 Rhodococcus erythropolis SQ1 34.3 62.1 303 3-ketosteroid dehydrogenase kstD1 2879 6379 2786355 2785651 705 sp.ALSR_BACSU Bacillus subtilis 168 alsR 37.1 69.0 232 transcriptional regulator, LysR family 2880 6380 2787782 2788594 813 pir:C70982 Mycobacterium tuberculosis 28.4 52.9 278 hypothetical protein H37Rv Rv3298c lpqC 2881 6381 2789399 2788587 813 pir:C69862 Bacillus subtilis 168 ykrA 26.7 55.6 288 hypothetical protein 2882 6382 2789935 2789477 459 2883 6383 2790152 2790550 399 pir:A45264 Oryctolagus cuniculus kidney 28.6 50.7 140 hypothetical protein cortex rBAT 2884 6384 2790946 2792448 1503 pir:B70798 Mycobacterium tuberculosis 36.0 64.0 464 hypothetical membrane protein H37Rv Rv3737 2885 6385 2792531 2792857 327 pir:S41307 Streptomyces griseus hrdB 32.3 50.3 155 transcription initiation factor sigma 2886 6386 2792873 2794327 1455 sp:TPS1_SCHPO Schizosaccharomyces pombe 38.8 66.7 487 trehalose-6-phosphate synthase tps1 2887 6387 2794300 2794812 513 2888 6388 2794870 2795637 768 sp:OTSB_ECOLI Escherichia coli K12 otsB 27.4 57.6 245 trehalose-phosphatase 2889 6389 2796749 2795676 1074 sp:CCPA_BACME Bacillus megaterium ccpA 24.7 60.2 344 glucose-resistance amylase regulator 2890 6390 2796865 2797806 942 sp:ZNUA_HAEIN Haemophilus influenzae Rd 22.4 46.7 353 high-affinity zinc uptake system HI0119 znuA protein 2891 6391 2797820 2798509 690 gp:AF121672_2 Staphylococcus aureus 8325-4 31.4 63.2 223 ABC transporter mreA 2892 6392 2798837 2799391 555 pir:E70507 Mycobacterium tuberculosis 60.0 87.4 135 hypothetical membrane protein H37Rv Rv2060 2893 6393 2799535 2801034 1500 pir:A69426 Archaeoglobus fulgidus 23.4 52.5 303 transposase (ISA0963-5) 2894 6394 2801113 2801313 201 2895 6395 2803246 2801558 1689 gp:AF096929_2 Rhodococcus erythropolis SQ1 32.1 62.0 561 3-ketosteroid dehydrogenase kstD1 2896 6396 2803996 2803250 747 2897 6397 2804691 2804074 618 pir:B72359 Thermotoga maritima MSB8 34.3 56.4 204 lipopolysaccharide biosynthesis bplA protein or oxidoreductase or dehydrogenase 2898 6398 2805110 2804676 435 sp:MI2D_BACSU Bacillus subtilis 168 idh or iolG 35.2 69.5 128 dehydrogenase or myo-inositol 2- dehydrogenase 2899 6399 2805967 2805113 855 sp:SHIA_ECOLI Escherichia coli K12 shiA 30.5 67.5 292 shikimate transport protein 2900 6400 2806441 2806016 426 sp:SHIA_ECOLI Escherichia coli K12 shiA 43.1 80.8 130 shikimate transport protein 2901 6401 2807252 2806599 654 gp:SC5A7_19 Streptomyces coelicolor A3(2) 32.6 55.7 212 transcriptional regulator SC5A7.19c 2902 6402 2808364 2807426 939 sp:PT56_YEAST Saccharomyces cerevisiae 22.8 47.3 334 ribosomal RNA ribose methylase or YOR201C PET56 tRNA/rRNA methyltransferase 2903 6403 2809778 2808399 1380 sp:SYC_ECOLI Escherichia coli K12 cysS 42.2 68.8 464 cysteinyl-tRNA synthetase 2904 6404 2811806 2809824 1983 prf:2511335C Lactococcus lactis sacB 47.0 77.0 668 PTS system, enzyme II sucrose protein (sucrose-specific IIABC component) 2905 6405 2813258 2811960 1299 gp:AF205034_4 Clostridium acetobutylicum 35.3 56.9 473 sucrose 6-phosphate hydrolase or ATCC 824 scrB sucrase 2906 6406 2814037 2813279 759 sp:NAGB_ECOLI Escherichia coli K12 nagB 38.3 69.4 248 glucosamine-6-phosphate isomerase 2907 6407 2815232 2814081 1152 sp:NAGA_VIBFU Vibrio furnissii SR1514 manD 30.2 60.3 368 N-acetylglucosamine-6-phosphate deacetylase 2908 6408 2815458 2816393 936 sp:DAPA_ECOLI Escherichia coli K12 dapA 28.2 62.1 298 dihydrodipicolinate synthase 2909 6409 2816409 2817317 909 sp:GLK_STRCO Streptomyces coelicolor A3(2) 28.7 57.6 321 glucokinase SC6E10.20c glk 2910 6410 2817363 2818058 696 prf:2516292A Clostridium perfringens NCTC 36.4 68.6 220 N-acetylmannosamine-6-phosphate 8798 nanE epimerase 2911 6411 2818313 2818137 177 2912 6412 2819564 2818350 1215 sp:NANH_MICVI Micromonospora viridifaciens 24.8 50.3 439 sialidase precursor ATCC 31146 nadA 2913 6413 2820285 2819557 729 gp:AF181498_1 Rhizobium etli ansR 26.6 57.2 222 L-asparagine permease operon repressor 2914 6414 2820584 2822191 1608 gp:BFU64514_1 Bacillus firmus OF4 dppA 22.5 51.4 560 dipeptide transporter protein or heme-binding protein 2915 6415 2822387 2823337 951 sp:DPPB_BACFI Bacillus firmus OF4 dappB 31.9 64.3 342 dipeptide transport system permease protein 2916 6416 2824274 2825341 1068 sp:OPPD_BACSU Bacillus subtilis 168 oppD 46.5 78.3 314 oligopeptide transport ATP-binding protein 2917 6417 2825341 2826156 816 sp:OPPF_LACLA Lactococcus lactis oppF 43.4 78.7 258 oligopeptide transport ATP-binding protein 2918 6418 2826835 2826215 621 sp:RHTB_ECOLI Escherichia coli K12 rhtB 28.5 62.7 193 homoserine/homoserin lactone efflux protein or lysE type translocator 2919 6419 2826922 2827404 483 prf:2309303A Bradyrhizobium japonicum lrp 31.0 66.2 142 leucine-responsive regulatory protein 2920 6420 2827817 2827458 360 2921 6421 2828383 2827904 480 pir:C70607 Mycobacterium tuberculosis 55.9 86.2 152 hypothetical protein H37Rv Rv3581c 2922 6422 2829146 2828379 768 sp:Y18T_MYCTU Mycobacterium tuberculosis 46.4 71.5 235 hypothetical protein H37Rv Rv3582c 2923 6423 2829749 2829156 594 pir:H70803 Mycobacterium tuberculosis 73.3 91.1 157 transcription factor H37Rv Rv3583c 2924 6424 2830057 2830779 723 prf:2214304A Mycobacterium tuberculosis 43.5 70.0 223 two-component system response H37Rv Rv3246c mtrA regulator 2925 6425 2830779 2831894 1116 sp:BAES_ECOLI Escherichia coli K12 baeS 29.3 67.7 341 two-component system sensor histidine kinase 2926 6426 2832085 2832666 582 2927 6427 2832790 2834181 1392 sp:RADA_ECOLI Escherichia coli K12 radA 41.5 74.3 463 DNA repair protein RadA 2928 6428 2834188 2835285 1098 sp:YACK_BACSU Bacillus subtilis 168 yacK 40.3 73.3 345 hypothetical protein 2929 6429 2835969 2835283 687 pir:D70804 Mycobacterium tuberculosis 29.4 53.3 231 hypothetical protein H37Rv Rv3587c 2930 6430 2837499 2836048 1452 gp:PPU96338_1 Pseudomonas putida NCIMB 59.5 85.1 471 p-hydroxybenzaldehyde 9866 plasmid pRA4000 dehydrogenase 2931 6431 2837737 2837591 147 2932 6432 2838576 2837956 621 pir:T08204 Chlamydomonas reinhardtii ca1 36.7 66.2 210 mitochondrial carbonate dehydratase beta 2933 6433 2838643 2839521 879 gp:AF121797_1 Streptomyces antibioticus IMRU 48.4 70.7 283 A/G-specific adenine glycosylase 3720 mutY 2934 6434 2839562 2840716 1155 2935 6435 2841063 2840758 306 2936 6436 2841075 2841848 774 gp:AB009078_1 Brevibacterium saccharolyticum 99.2 99.6 258 L-2.3-butanediol dehydrogenase 2937 6437 2842130 2842453 324 2938 6438 2842493 2843233 741 2939 6439 2843405 2843716 312 2940 6440 2843722 2843432 291 pir:E70552 Mycobacterium tuberculosis 48.5 69.1 97 hypothetical protein H37Rv Rv3592 2941 6441 2845139 2845558 420 GSP:Y29188 Pseudomonas aeruginosa 57.0 63.0 99 virulence factor 0RF24222 2942 6442 2845889 2846101 213 GSP:Y29193 Pseudomonas aeruginosa 54.0 55.0 72 virulence factor 0RF25110 2943 6443 2846186 2846506 321 GSP:Y29193 Pseudomonas aeruginosa 74.0 75.0 55 virulence factor ORF25110 2944 6444 2846940 2844166 2775 sp:MECB_BACSU Bacillus subtilis 168 mecB 58.5 86.2 832 ClpC adenosine triphosphatase/ ATP-binding proteinase 2945 6445 2847229 2848659 1431 gp:AB035643_1 Bacillus cereus ts-4 impdh 37.1 70.2 469 inosine monophosphate dehydrogenase 2946 6446 2848769 2849779 1011 pir:JC6117 Rhodococcus rhodochrous nitR 24.7 62.7 316 transcription factor 2947 6447 2850031 2851815 1785 sp:PH2M_TRICU Trichosporon cutaneum ATCC 33.5 60.9 680 phenol 2-monooxygenase 46490 2948 6448 2852017 2853732 1716 2949 6449 2853769 2855709 1941 2950 6450 2855795 2857516 1722 2951 6451 2859044 2859205 162 2952 6452 2859055 2857613 1443 gp:AF237667_1 Corynebacterium glutamicum 100.0 100.0 481 lincomycin resistance protein lmrB 2953 6453 2860145 2859195 951 pir:G70807 Mycobacterium tuberculosis 26.7 55.8 240 hypothetical protein H37Rv Rv3517 2954 6454 2862082 2860505 1578 gp:AB012100_1 Bacillus stearothermophilus lysS 41.7 71.2 511 lysyl-tRNA synthetase 2955 6455 2862929 2862132 798 gp:CGPAN_2 Corynebacterium glutamicum 29.9 52.6 268 pantoate-beta-alanine ligase ATCC 13032 panC 2956 6456 2863621 2862929 693 2957 6457 2864421 2863624 798 2958 6458 2864848 2864384 465 gp:MLCB2548_4 Mycobacterium leprae 29.0 69.6 138 hypothetical membrane protein MLCB2548.04c 2959 6459 2865343 2864867 477 sp:HPPK_METEX Methylobacterium extorquens 42.4 69.0 158 2-amino-4-hydroxy-6- AM1 folK hydroxymethyldihydropteridine pyrophosphokinase 2960 6460 2865735 2865346 390 sp:FOLB_BACSU Bacillus subtilis 168 folB 38.1 69.5 118 dihydroneopterin aldolase 2961 6461 2866567 2865731 837 gp:AB028656_1 Mycobacterium leprae folP 51.5 75.0 268 dihydropteroate synthase 2962 6462 2867173 2866586 588 sp:GCH1_BACSU Bacillus subtilis 168 mtrA 60.6 86.2 188 GTP cyclohydrolase I 2963 6463 2867471 2868385 915 2964 6464 2869748 2867169 2580 56.0 69.0 782 cell division protein FtsH 2965 6465 2870444 2869863 582 gp:AF008931_1 Salmonella typhimurium GP660 51.5 83.0 165 hypoxanthine hprt phosphoribosyltransferase 2966 6466 2871389 2870499 891 sp:YZC5_MYCTU Mycobacterium tuberculosis 41.0 66.8 310 cell cycle protein MesJ or cytosine H37Rv Rv3625c deaminase-related protein 2967 6467 2872677 2871445 1233 sp:DAC_ACTSP Actinomadura sp. R39 dac 27.2 51.4 459 D-alanyl-D-alanine carboxypeptidase 2968 6468 2872926 2873399 474 sp:IPYR_ECOLI Escherichia coli K12 ppa 49.7 73.6 159 inorganic pyrophosphatase 2969 6469 2873611 2873393 219 2970 6470 2875443 2873905 1539 pir:H70886 Mycobacterium tuberculosis 56.0 80.7 507 spermidine synthase H37Rv speE 2971 6471 2875832 2875434 399 sp:YOB1_MYCTU Mycobacterium tuberculosis 38.6 86.4 132 hypothetical membrane protein H37Rv Rv2600 2972 6472 2876280 2875870 411 sp:YOB2_MYCTU Mycobacterium tuberculosis 36.8 63.2 144 hypothetical protein H37Rv Rv2599 2973 6473 2876777 2876280 498 sp:YOB3_MYCTU Mycobacterium tuberculosis 36.4 60.1 173 hypothetical protein H37Rv Rv2598 2974 6474 2877385 2876777 609 sp:YOB4_MYCTU Mycobacterium tuberculosis 44.6 72.3 202 hypothetical protein H37Rv Rv2597 2975 6475 2877703 2877455 249 sp:PTBA_BACSU Bacillus subtilis 168 bglP 30.3 59.6 89 PTS system, beta-glucosides- permease II ABC component 2976 6476 2877858 2877595 264 2977 6477 2879710 2878478 1233 gp:AB017795_2 Nocardioides sp. KP7 phdD 38.0 69.6 411 ferredoxin reductase 2978 6478 2879965 2880252 288 gp:SCH69_9 Streptomyces coelicolor A3(2) 46.4 73.2 97 hypothetical protein SCH69.09c 2979 6479 2880544 2880987 444 prf:2516298U Burkholderia pseudomallei ORF 26.7 59.3 135 bacterial regulatory protein, marR E family 2980 6480 2880998 2884882 3885 prf:2413335A Streptomyces roseosporus cpsB 28.4 51.6 1241 peptide synthase 2981 6481 2883304 2881844 1461 2982 6482 2886497 2884935 1563 prf:2310295A Escherichia coli K12 padA 35.0 63.7 488 phenylacetaldehyde dehydrogenase 2983 6483 2887833 2886916 918 gp:CJ11168X2_254 Campylobacter jejuni Cj0604 57.3 79.7 241 hypothetical protein 2984 6484 2890185 2890346 162 GP:MSGTCWPA_1 Mycobacterium tuberculosis 62.0 63.0 54 hypothetical protein 2985 6485 2890377 2890553 177 GP:MSGTCWPA_1 Mycobacterium tuberculosis 74.0 80.0 31 hypothetical protein 2986 6486 2890540 2888897 1644 gsp:R94368 Brevibacterium flavum MJ-233 99.5 100.0 548 heat shock protein or chaperon or groEL protein 2987 6487 2890930 2890751 180 2988 6488 2892138 2890930 1209 2989 6489 2893100 2892138 963 2990 6490 2895085 2893100 1986 2991 6491 2897525 2895072 2454 2992 6492 2900326 2897528 2799 2993 6493 2903920 2900330 3591 prf:2309326A Homo sapiens MUC5B 21.7 42.3 1236 hypothetical protein 2994 6494 2906738 2903964 2775 2995 6495 2907250 2906639 612 2996 6496 2907515 2908885 1371 pir:G70870 Mycobacterium tuberculosis 37.1 68.0 447 peptidase H37Rv Rv2522c 2997 6497 2909210 2909788 579 2998 6498 2909830 2909231 600 2999 6499 2910172 2913228 3057 prf:2504285B Staphylococcus aureus mnhA 35.6 68.3 797 Na+/H+ antiporter or multiple resistance and pH regulation related protein A or NADH dehydrogenase 3000 6500 2913235 2913723 489 gp:AF097740_3 Bacillus firmus OF4 mrpC 44.2 81.7 104 Na+/H+ antiporter or multiple resistance and pH regulation related protein C or cation transport system protein 3001 6501 2913749 2915416 1668 gp:AF097740_4 Bacillus firmus OF4 mrpD 35.2 72.1 523 Na+/H+ antiporter or multiple resistance and pH regulation related protein D 3002 6502 2915482 2915922 441 gp:AF097740_5 Bacillus firmus OF4 mrpE 26.7 60.9 161 Na+/H+ antiporter or multiple resistance and pH regulation related protein E 3003 6503 2915929 2916201 273 prf:2416476G Rhizobium meliloti phaF 32.5 66.2 77 K+ efflux system or multiple resistance and pH regulation related protein F 3004 6504 2916205 2916582 378 prf:2504285H Staphylococcus aureus mnhG 25.6 63.6 121 Na+/H+ antiporter or multiple resistance and pH regulation related protein G 3005 6505 2917617 2917024 594 pir:D70594 Mycobacterium tuberculosis 24.7 54.5 178 hypothetical protein H37Rv lipV 3006 6506 2918757 2917630 1128 sp:YBDK_ECOLI Escherichia coli K12 ybdK 27.0 61.7 334 hypothetical protein 3007 6507 2919481 2918819 663 3008 6508 2919715 2920293 579 sp:DEF_BACSU Bacillus subtilis 168 def 37.5 60.9 184 polypeptide deformylase 3009 6509 2919741 2919490 252 pir:D70631 Mycobacterium tuberculosis 47.9 70.4 71 hypothetical protein H37Rv Rv0430 3010 6510 2920286 2921290 1005 pir:B70631 Mycobacterium tuberculosis 31.3 54.2 339 acetyltransferase (GNAT) family or H37Rv Rv0428c N terminal acetylating enzyme 3011 6511 2920476 2919808 669 3012 6512 2920849 2920220 630 3013 6513 2921320 2922108 789 gp:AF108767_1 Salmonella typhimurium LT2 30.8 59.9 31 exodeoxyribonuclease III or xthA exonuclease 3014 6514 2922118 2923617 1500 gp:BFU88888_2 Bacillus firmus OF4 cls 27.9 62.0 513 cardiolipin synthase 3015 6515 2924191 2924844 654 3016 6516 2925147 2923954 1194 sp:BCR_ECOLI Escherichia coli K12 bcr 31.6 67.2 393 membrane transport protein or bicyclomycin resistance protein 3017 6517 2925541 2926704 1164 gp:VCAJ10968_1 Vibrio cholerae JS1569 nptA 28.5 68.9 382 sodium dependent phosphate pump 3018 6518 2927546 2926707 840 sp:PHZC_PSEAR Pseudomonas aureofaciens 30- 38.8 56.4 289 phenazine biosynthesis protein 84 phzC 3019 6519 2928283 2927651 633 3020 6520 2928318 2927551 768 gp:SCE8_16 Streptomyces coelicolor A3(2) 24.3 60.8 255 ABC transporter SCE8.16c 3021 6521 2929237 2928302 936 sp:BCRA_BACLI Bacillus licheniformis ATCC 36.9 66.3 309 ABC transporter ATP-binding protein 9945A bcrA 3022 6522 2929756 2929256 501 pir:C70629 Mycobacterium tuberculosis 47.6 68.5 168 mutator mutT protein H37Rv Rv0413 3023 6523 2929951 2931336 1386 pir:B70629 Mycobacterium tuberculosis 35.0 70.2 423 hypothetical membrane protein H37Rv Rv0412c 3024 6524 2931340 2932371 1032 sp:GLNH_BACST Bacillus stearothermophilus 31.5 64.8 270 glutamine-binding protein precursor NUB36 glnH 3025 6525 2932577 2934829 2253 pir:H70628 Mycobacterium tuberculosis 41.2 63.5 805 serine/threonine kinase H37Rv Rv0410c pknG 3026 6526 2933398 2932652 747 3027 6527 2938403 2939767 1365 sp:ADRO_BOVIN Bos taurus 37.2 67.8 457 ferredoxin/ferredoxin-NADP reductase 3028 6528 2939907 2940452 546 sp:ELAA_ECOLI Escherichia coli K12 elaA 34.0 60.3 156 acetyltransferase (GNAT) family 3029 6529 2941508 2940447 1062 3030 6530 2942500 2941472 1029 3031 6531 2943007 2942609 399 3032 6532 2944205 2943012 1194 sp:PURT_BACSU Bacillus subtilis 168 purT 59.1 82.6 379 phosphoribosylglycinamide formyltransferase 3033 6533 2946526 2945639 888 3034 6534 2947591 2946698 894 pir:S60890 Corynebacterium glutamicum 77.6 90.9 295 insertion element (IS3 related) orf2 3035 6535 2947886 2947620 267 pir:S60889 Corynebacterium glutamicum 67.4 84.3 89 insertion element (IS3 related) orf1 3036 6536 2949188 2948049 1140 gp:AB016841_1 Streptomyces thermoviolaceus 22.4 51.3 349 two-component system sensor opc-520 chiS histidine kinase 3037 6537 2949882 2949265 618 sp:DEGU_BACBR Bacillus brevis ALK36 degU 31.7 65.6 218 transcriptional regulator 3038 6538 2950207 2950431 225 3039 6539 2951723 2950434 1290 gp:AB003160_1 Corynebacterium 89.7 95.3 427 adenylosuccinate synthetase ammoniagenes purA 3040 6540 2951933 2952691 759 pir:G70575 Mycobacterium tuberculosis 34.3 59.3 204 hypothetical protein H37Rv Rv0358 3041 6541 2952709 2952972 264 3042 6542 2954141 2952975 1167 sp:YFDA_CORGL Corynebacterium glutamicum 100.0 100.0 359 hypothetical membrane protein AS019 ATCC 13059 ORF3 3043 6543 2955272 2954241 1032 pir:S09283 Corynebacterium glutamicum 99.7 100.0 344 fructose-bisphosphate aldolase AS019 ATCC 13059 fda 3044 6544 2956473 2955523 951 gp:CGFDA_1 Corynebacterium glutamicum 100.0 100.0 304 hypothetical protein AS019 ATCC 13059 ORF1 3045 6545 2957447 2956830 618 pir:G70833 Mycobacterium tuberculosis 76.9 91.2 182 methyltransferase H37Rv Rv0380c 3046 6546 2958036 2957485 552 gp:AF058713_1 Pyrococcus abyssi pyrE 39.1 65.5 174 orotate phosphoribosyltransferase 3047 6547 2959110 2958139 972 pir:B70834 Mycobacterium tuberculosis 27.6 60.0 250 hypothetical protein H37Rv Rv0383c 3048 6548 2960371 2959520 852 sp:THTM_HUMAN Homo sapiens mpsT 29.6 56.1 294 3-mercaptopyruvate sulfurtransferase 3049 6549 2961187 2960468 720 3050 6550 2963008 2962730 279 3051 6551 2963596 2963198 399 3052 6552 2964258 2964434 177 GSP:Y29188 Pseudomonas aeruginosa 76.0 82.0 59 virulence factor ORF24222 3053 6553 2965076 2965837 762 GSP:Y29182 Pseudomonas aeruginosa 38.0 55.0 200 virulence factor ORF23228 3054 6554 2965188 2965583 396 GSP:Y29193 Pseudomonas aeruginosa 62.0 63.0 132 virulence factor ORF25110 3055 6555 2967804 2966458 1347 pir:S76683 Synechocystis sp. PCC6803 24.7 54.8 489 sodium/glutamate symport carrier slr0625 protein 3056 6556 2968403 2968789 387 sp:CADF_STAAU Staphylococcus aureus cadC 37.0 71.3 108 cadmium resistance protein 3057 6557 2968951 2969808 858 pir:H75109 Pyrococcus abyssi Orsay 23.7 63.3 283 cation efflux system protein PAB0462 (zinc/cadmium) 3058 6558 2969834 2971003 1170 gp:AB010439_1 Rhodococcus rhodochrous 22.5 45.4 476 monooxygenase or oxidoreductase IFO3338 or steroid monooxygenase 3059 6559 2971017 2972057 1041 sp:LUXA_KRYAS Kryptophanaron alfredi symbiont 21.1 47.4 399 alkanal monooxygenase alpha chain luxA 3060 6560 2972099 2971338 762 3061 6561 2973205 2972060 1146 sp:METB_ECOLI Escherichia coli K12 metB 36.5 62.4 375 cystathionine gamma-lyase 3062 6562 2973796 2973230 567 gp:SC1A2_11 Streptomyces coelicolor A3(2) 40.2 67.9 184 bacterial regulatory protein, lacl SC1A2.11 family 3063 6563 2973961 2974200 240 gp:SCE20_34 Streptomyces coelicolor A3(2) 49.4 65.2 89 rifampin ADP-ribosyl transferase SCE20.34c arr 3064 6564 2974200 2974382 183 gp:SCE20_34 Streptomyces coelicolor A3(2) 73.2 87.5 56 rifampin ADP-ribosyl transferase SCE20.34c arr 3065 6565 2974467 2975591 1125 pir:E70812 Mycobacterium tuberculosis 30.5 56.2 361 hypothetical protein H37Rv Rv0837c 3066 6566 2975629 2976360 732 pir:D70812 Mycobacterium tuberculosis 33.8 64.7 204 hypothetical protein H37Rv Rv0836c 3067 6567 2976596 2977774 1179 pir:D70834 Mycobacterium tuberculosis 31.9 60.6 386 oxidoreductase H37Rv Rv0385 3068 6568 2978644 2977847 798 pir:B69109 Methanobacterium 32.0 67.3 275 N-carbamoyl-D-amino acid thermoautotrophicum Delta H amidohydrolase MTH1811 3069 6569 2978737 2978979 243 3070 6570 2978982 2980115 1134 gp:SC4A7_3 Streptomyces coelicolor A3 (2) 28.0 55.4 289 hypothetical protein SC4A7.03 3071 6571 2980887 2981216 330 GP:ABCARRA_2 Azospirillum brasilense carR 38.0 44.0 108 novel two-component regulatory system 3072 6572 2981698 2980181 1518 prf.2104333D Rhodococcus erythropolis thcA 69.6 90.3 507 aldehyde dehydrogenase 3073 6573 2982460 2982023 438 gp:SAU43299_2 Streptomyces albus G hspR 47.4 70.4 135 heat shock transcription regulator 3074 6574 2983679 2982495 1185 sp:DNAJ_MYCTU Mycobacterium tuberculosis 56.7 80.1 397 heat shock protein dnaJ H37Rv RV0352 dnaJ 3075 6575 2984522 2983887 636 sp:GRPE_STRCO Streptomyces coelicolor grpE 38.7 66.5 212 nucleotide exchange factor grpE protein bound to the ATPase domain of the molecular chaperone DnaK 3076 6576 2986397 2984544 1854 gsp:R94587 Brevibacterium flavum MJ-233 99.8 99.8 618 heat shock protein dnaK dnaK 3077 6577 2986833 2988164 1332 gp:SCF6_8 Streptomyces coelicolor A3(2) 42.6 79.0 338 hypothetical membrane protein SCF6.09 3078 6578 2988846 2988214 633 sp:PFS_HELPY Helicobacter pylori HP0089 mtn 27.2 60.0 195 5′-methylthioadenosine nucleosidase and S- adenosylhomocysteine nucleosidase 3079 6579 2990045 2988846 1200 3080 6580 2991718 2992602 885 3081 6581 2993286 2989954 3333 sp:CUT3_SCHPO Schizosaccharomyces pombe 18.9 48.4 1311 chromosome segregation protein cut3 3082 6582 2993921 2993286 636 3083 6583 2995405 2993921 1485 3084 6584 2996781 2995747 1035 sp:ADH2_BACST Bacillus stearothermophilus 50.0 81.7 334 alcohol dehydrogenase DSM 2334 adh 3085 6585 2997151 2997366 216 3086 6586 2997687 2997481 207 3087 6587 2997688 2997876 189 3088 6588 2998223 2997963 261 3089 6589 2999454 2998528 927 pir:F69997 Bacillus subtilis ytnM 43.5 70.1 301 hypothetical membrane protein 3090 6590 3000200 2999478 723 gp:SC7A8_10 Streptomyces coelicolor A3(2) 32.5 53.2 252 hypothetical protein SC7A8.10c 3091 6591 3001512 3002426 915 3092 6592 3001539 3000241 1299 sp:CYSN_ECOLI Escherichia coli K12 cysN 47.3 78.3 414 sulfate adenylyltransferase, subunit 1 3093 6593 3002453 3001542 912 sp:CYSD_ECOLI Escherichia coli K12 cysD 46.1 70.1 308 sulfate adenylyltransferase small chain 3094 6594 3003145 3002453 693 sp:CYH1_BACSU Bacillus subtilis cysH 39.2 64.2 212 phosphoadenosine phosphosulfate reductase 3095 6595 3005162 3003480 1683 sp:NIR_SYNP7 Synechococcus sp. PCC 7942 34.5 65.5 502 ferredoxin-nitrate reductase 3096 6596 3005545 3006915 1371 sp:ADRO_YEAST Saccharomyces cerevisiae 30.8 61.4 487 ferredoxin/ferredoxin-NADP FL200 arh1 reductase 3097 6597 3007294 3008376 1083 prf:2420294J Homo sapiens hypE 32.6 59.7 144 huntingtin interactor 3098 6598 3008689 3008453 237 3099 6599 3008770 3009303 534 3100 6600 3009162 3008749 414 sp:PHNB_ECOLI Escherichia coli K12 phnB 26.8 59.9 142 alkylphosphonate uptake protein and C-P lyase activity 3101 6601 3009242 3009607 366 gp:SCE68_10 Streptomyces coelicolor A3(2) 50.0 66.3 80 hypothetical protein SCE68.10 3102 6602 3010231 3009710 522 gp:PPAMOA_1 Pseudomonas putida DSMZ ID 39.1 76.4 161 ammonia monooxygenase 88-260 amoA 3103 6603 3010659 3010979 321 3104 6604 3010926 3010441 486 3105 6605 3010989 3011273 285 SP:YTZ3_AGRVI Agrobacterium vitis ORFZ3 41.0 58.0 68 hypothetical protein 3106 6606 3011805 3011242 564 3107 6607 3012809 3011808 1002 sp:YGB7_ALCEU Alcaligenes eutrophus H16 26.1 57.9 337 hypothetical protein ORF7 3108 6608 3013798 3013106 693 gp:HIU68399_3 Haemophilus influenzae hmcB 35.7 64.8 199 ABC transporter 3109 6609 3014550 3013837 714 gp:HIU68399_3 Haemophilus influenzae hmcB 39.3 73.0 211 ABC transporter 3110 6610 3014616 3015824 1209 pir:A69778 Bacillus subtilis ydeG 30.8 67.8 416 metabolite transport protein homolog 3111 6611 3015469 3014648 822 3112 6612 3016238 3016924 687 3113 6613 3017149 3015827 1323 sp:DAPE _ECOLI Escherichia coli K12 msgB 21.5 48.5 466 succinyl-diaminopimelate desuccinylase 3114 6614 3017316 3019220 1905 3115 6615 3017539 3018312 774 3116 6616 3018181 3017420 762 3117 6617 3019076 3018123 954 GPU:DCA297422—1 Daucus carota 33.0 46.0 114 dehydrin-like protein 3118 6618 3020609 3019542 1068 sp:MALK_ECOLI Escherichia coli K12 malK 24.9 50.1 373 maltose/maltodextrin transport ATP- binding protein 3119 6619 3021202 3020561 642 3120 6620 3021825 3021208 618 gp:AF036485_6 Lactococcus lactis Plasmid 30.2 67.6 179 cobalt transport protein pNZ4000 Orf-200 cbiM 3121 6621 3022928 3022113 816 sp:FRP_VIBHA Vibrio harveyi MAV frp 37.2 71.4 231 NADPH-flavin oxidoreductase 3122 6622 3023900 3022998 903 sp:IUNH_CRIFA Crithidia fasciculata iunH 28.4 59.3 317 inosine-uridine preferring nucleoside hydrolase 3123 6623 3024379 3025353 975 gp:SCE20_8 Streptomyces coelicolor A3(2) 31.2 59.4 276 hypothetical membrane protein SCE20.08c 3124 6624 3025552 3026139 588 sp:3MG1_ECOLI Escherichia coli K12 tag 50.3 78.8 179 DNA-3-methyladenine glycosylase 3125 6625 3027299 3026142 1158 sp:HMPA_ALCEU Alcaligenes eutrophus H16 fhp 33.5 63.8 406 flavohemoprotein 3126 6626 3027561 3028163 603 3127 6627 3028268 3028891 624 gp:SCO276673_18 Streptomyces coelicolor A3(2) 34.8 63.8 210 oxidoreductase mmyQ 3128 6628 3028878 3029033 156 3129 6629 3029474 3028884 591 sp:BGLG_ECOLI Escherichia coli K12 bglC 28.1 69.3 192 transcription antiterminator or beta- glucoside positive regulatory protein 3130 6630 3029504 3029782 279 3131 6631 3030061 3029702 360 sp:ABGA _CLOLO Clostridium longisporum B6405 43.7 59.9 167 6-phospho-beta-glucosidase abgA 3132 6632 3030155 3030535 381 3133 6633 3030340 3030101 240 sp:ABGA _CLOLO Clostridium longisporum B6405 43.9 78.8 66 6-phospho-beta-glucosidase abgA 3134 6634 3030723 3031979 1257 gp:L78665_2 Methylobacillus flagellatus aat 53.7 80.9 402 aspartate aminotransferase 3135 6635 3032647 3032348 300 3136 6636 3032661 3033863 1203 gp:AF189147_1 Corynebacterium glutamicum 100.0 100.0 401 transposase (ISCg2) ATCC 13032 tnp 3137 6637 3034181 3035437 1257 gp:SCQ11_10 Streptomyces coelicolor A3(2) 33.6 70.2 399 hypothetical membrane protein SCQ11.10c 3138 6638 3034287 3034105 183 3139 6639 3036756 3035440 1317 prf:2422381B Sinorhizobium meliloti rkpK 40.5 72.2 442 UDP-glucose dehydrogenase 3140 6640 3037411 3036845 567 sp:DCD_ECOLI Escherichia coli K12 dcd 43.6 72.3 188 deoxycytidine triphosphate deaminase 3141 6641 3037675 3037911 237 3142 6642 3038172 3038942 771 gp:SCC75A _16 Streptomyces coelicolor A3(2) 30.6 59.4 229 hypothetical protein SCC75A.16c 3143 6643 3040681 3038993 1689 3144 6644 3041932 3040748 1185 gp:AB008771_1 Streptomyces thermoviolaceus 28.5 58.1 410 beta-N-Acetylglucosaminidase nagA 3145 6645 3041994 3042437 444 3146 6646 3042503 3042703 201 3147 6647 3042660 3045788 3129 gp:MLCB1883_7 Mycobacterium leprae 29.6 49.4 1416 hypothetical protein MLCB1883.13c 3148 6648 3043642 3043022 621 3149 6649 3045796 3045990 195 3150 6650 3047146 3048048 903 gp:MLCB1883_4 Mycobacterium leprae 24.8 47.1 363 hypothetical membrane protein MLCB1883.05c 3151 6651 3047189 3046122 1068 pir:JC4001 Streptomyces sp. acyA 27.7 51.0 408 acyltransferase or macrolide 3-O- acyltransferase 3152 6652 3047904 3047197 708 3153 6653 3048058 3049479 1422 gp:MLCB1883_3 Mycobacterium leprae 31.2 54.8 529 hypothetical membrane protein MLCB1883.04c 3154 6654 3050522 3051190 669 3155 6655 3050592 3049456 1137 pir:G70961 Mycobacterium tuberculosis 53.4 79.1 369 hexosyltransferase H37Rv Rv0225 3156 6656 3051194 3051964 771 pir:F70961 Mycobacterium tuberculosis 58.6 73.3 251 methyl transferase H37Rv Rv0224c 3157 6657 3053891 3052062 1830 sp:PPCK _NEOFR Neocallimastix frontalis pepck 54.7 78.5 601 phosphoenolpyruvate carboxykinase (GTP) 3158 6658 3054759 3055769 1011 pir:E75125 Pyrococcus abyssi Orsay 24.4 52.7 332 C4-dicarboxylate transporter PAB2393 3159 6659 3055867 3056631 765 sp:YGGH_ECOLI Escherichia coli K12 yggH 35.7 67.2 241 hypothetical protein 3160 6660 3056613 3057317 705 pir:E70959 Mycobacterium tuberculosis 69.1 85.0 207 hypothetical protein H37Rv Rv0207c 3161 6661 3057328 3059643 2316 pir:C70839 Mycobacterium tuberculosis 42.3 72.3 768 mebrane transport protein H37Rv Rv0206c mmpL3 3162 6662 3059517 3058096 1422 3163 6663 3059651 3060733 1083 pir:A70839 Mycobacterium tuberculosis 29.1 62.9 364 hypothetical membrane protein H37Rv Rv0204c 3164 6664 3060733 3061095 363 pir:H70633 Mycobacterium tuberculosis 34.3 69.4 108 hypothetical membrane protein H37Rv Rv0401 3165 6665 3062927 3061380 1548 gp:AF113605_1 Streptomyces coelicolor A3(2) 49.7 76.9 523 propionyl-CoA carboxylase complex pccB B subunit 3166 6666 3067780 3062951 4830 sp:ERY1_SACER Streptomyces erythraeus eryA 30.2 54.2 1747 polyketide synthase 3167 6667 3069930 3068143 1788 prf:2310345A Mycobacterium bovis BCG 33.5 62.3 592 acyl-CoA synthase 3168 6668 3071140 3070214 927 pir:F70887 Mycobacterium tuberculosis 39.8 67.4 319 hypothetical protein H37Rv Rv3802c 3169 6669 3071644 3071147 498 3170 6670 3073620 3071650 1971 sp:CSP1_CORGL Corynebacterium glutamicum 98.6 99.5 657 major secreted protein PS1 protein (Brevibacterium flavum) ATCC precursor 17965 cop1 3171 6671 3074047 3075447 1401 3172 6672 3074075 3073857 219 3173 6673 3076562 3075540 1023 sp:A85C_MYCTU Mycobacterium tuberculosis 36.3 62.5 331 antigen 85-C ERDMANN RV0129C fbpC 3174 6674 3078772 3076715 2058 pir:A70888 Mycobacterium tuberculosis 37.5 61.2 667 hypothetical membrane protein H37Rv Rv3805c 3175 6675 3079848 3078853 996 sp:NOEC_AZOCA Azorhizobium caulinodans 27.1 51.5 295 nodulation protein ORS571 noeC 3176 6676 3080351 3079848 504 pir:C70888 Mycobacterium tuberculosis 51.2 75.0 168 hypothetical protein H37Rv Rv3807c 3177 6677 3082311 3080344 1968 pir:D70888 Mycobacterium tuberculosis 55.6 74.7 656 hypothetical protein H37Rv Rv3808c 3178 6678 3082467 3083960 1494 3179 6679 3084411 3083935 477 sp:BCRC_BACLI Bacillus licheniformis ATCC 28.2 56.5 170 phosphatidic acid phosphatase 9945A bcrC 3180 6680 3085200 3084424 777 3181 6681 3085727 3085218 510 3182 6682 3085747 3087048 1302 sp:FMO1_PIG Sus scrofa fmo1 24.4 50.4 377 dimethylaniline monooxygenase (N- oxide-forming) 3183 6683 3087665 3088276 612 3184 6684 3088303 3087101 1203 sp:GLF_ECOLI Escherichia coli K12 glf 43.2 72.9 377 UDP-galactopyranose mutase 3185 6685 3088616 3090664 2049 pir:G70520 Mycobacterium tuberculosis 29.6 47.8 659 hypothetical protein H37Rv Rv3811 csp 3186 6686 3092286 3090760 1527 sp:GLPK _PSEAE Pseudomonas aeruginosa 51.7 78.8 499 glycerol kinase ATCC 15692 glpK 3187 6687 3093175 3092342 834 pir:A70521 Mycobacterium tuberculosis 41.6 70.3 279 hypothetical protein H37Rv Rv3813c 3188 6688 3094050 3093175 876 pir:D70521 Mycobacterium tuberculosis 46.7 72.0 261 acyltransferase H37Rv Rv3816c 3189 6689 3095343 3094078 1266 gsp:W26465 Mycobacterium tuberculosis 70.2 87.6 419 seryl-tRNA synthetase H37Rv 3190 6690 3095574 3096287 714 sp:FARR _ECOLI Escherichia coli K12 farR 27.7 61.7 235 transcriptional regulator, GntR family or fatty acyl-responsive regulator 3191 6691 3096311 3097423 1113 pir:H70652 Mycobacterium tuberculosis 32.6 61.2 356 hypothetical protein H37Rv Rv3835 3192 6692 3097423 3097764 342 pir:A70653 Mycobacterium tuberculosis 46.0 79.7 113 hypothetical protein H37Rv Rv3836 3193 6693 3097878 3097780 99 3194 6694 3098572 3097904 669 gp:AMU73808_1 Amycolatopsis methanolica pgm 37.2 62.8 218 2,3-PDG dependent phosphoglycerate mutase 3195 6695 3098825 3099454 630 3196 6696 3099556 3100698 1143 prf:2501285A Mycobacterium smegmatis pzaA 27.4 50.9 460 nicotinamidase or pyrazinamidase 3197 6697 3100698 3101426 729 3198 6698 3101734 3102768 1035 gp:SC6G4_33 Streptomyces coelicolor A3(2) 31.6 57.1 380 transcriptional regulator SC6G4.33 3199 6699 3101863 3101744 120 3200 6700 3102630 3102079 552 3201 6701 3102894 3103763 870 3202 6702 3103926 3104252 327 pir:B26872 Streptomyces lavendulae 43.9 81.3 107 hypothetical protein ORF372 3203 6703 3104406 3105719 1314 sp:AMYH_YEAST Saccharomyces cerevisiae 28.7 55.3 432 glucan 1,4-alpha-glucosidase S288C YIR019C sta1 3204 6704 3106970 3106053 918 3205 6705 3107769 3106951 819 sp:GLPQ_BACSU Bacillus subtilis glpQ 29.0 54.1 259 glycerophosphoryl diester phosphodiesterase 3206 6706 3108131 3109519 1389 sp:GNTP_BACSU Bacillus subtilis gntP 37.3 71.9 456 gluconate permease 3207 6707 3109464 3108823 642 3208 6708 3109845 3110003 159 3209 6709 3112080 3110464 1617 sp:KPYK_CORGL Corynebacterium glutamicum 25.5 47.7 491 pyruvate kinase AS019 pyk 3210 6710 3113390 3112449 942 gsp:Y25997 Brevibacterium flavum lctA 99.7 99.7 314 L-lactate dehydrogenase 3211 6711 3113619 3115394 1776 pir:C70893 Mycobacterium tuberculosis 33.5 64.8 526 hypothetical protein H37Rv Rv1069c 3212 6712 3115407 3116042 636 gp:SC1C2_30 Streptomyces coelicolor A3(2) 32.1 58.5 224 hydrolase or haloacid SC1C2.30 dehalogenase-like hydrolase 3213 6713 3116079 3116621 543 gp:AF030288_1 Brevibacterium linens ORF1 39.9 67.6 188 efflux protein tmpA 3214 6714 3116640 3117332 693 sp:GLCC_ECOLI Escherichia coli K12 MG1655 27.6 57.0 221 transcription activator or glcC transcriptional regulator GntR family 3215 6715 3117336 3118121 786 pir:B70885 Mycobacterium tuberculosis 47.8 68.6 255 phosphoesterase H37Rv Rv2795c 3216 6716 3118284 3119582 1299 sp:SHIA_ECOLI Escherichia coli K12 shiA 37.9 74.4 422 shikimate transport protein 3217 6717 3119665 3120879 1215 prf:2219306A Neisseria meningitidis lldA 40.4 68.9 376 L-lactate dehydrogenase or FMN- dependent dehydrogenase 3218 6718 3120909 3121313 405 3219 6719 3121598 3121909 312 sp:RPC_BPPH1 Bacillus phage phi-105 ORF1 45.5 80.0 55 immunity repressor protein 3220 6720 3122129 3121992 138 3221 6721 3123222 3123932 711 3222 6722 3124172 3122556 1617 gp:CELY51B11A_1 Caenorhabditis elegans 29.5 51.3 569 phosphatase or reverse Y51B11A.1 transcriptase (RNA-dependent) 3223 6723 3124886 3124341 546 3224 6724 3125298 3124897 402 sp:ILL1_ARATH Arabidopsis thaliana ill 1 36.9 63.1 122 peptidase or IAA-amino acid hydrolase 3225 6725 3125343 3125492 150 3226 6726 3126145 3125495 651 sp:PMSR_ECOLI Escherichia coli B msrA 47.6 69.1 210 peptide methionine sulfoxide reductase 3227 6727 3126392 3126991 600 pir:I40858 Corynebacterium 82.3 92.7 164 superoxide dismutase (Fe/Mn) pseudodiphtheriticum sod 3228 6728 3128417 3127494 924 sp:GLTC_BACSU Bacillus subtilis gltC 32.5 65.8 292 transcriptional regulator 3229 6729 3128606 3129739 1134 gp:AF121000_10 Corynebacterium glutamicum 23.4 49.0 384 multidrug resistance transporter tetA 3230 6730 3129785 3131395 1611 3231 6731 3132920 3133030 111 3232 6732 3133028 3131508 1521 3233 6733 3133115 3133747 633 pir:G70654 Mycobacterium tuberculosis 33.8 64.8 216 hypothetical protein H37Rv Rv3850 3234 6734 3135268 3133778 1491 prf:2508244AB Streptomyces cyanogenus lanJ 27.3 59.3 447 membrane transport protein 3235 6735 3135297 3135752 456 sp:YXAD_BACSU Bacillus subtilis 168 yxaD 37.2 65.0 137 transcriptional regulator 3236 6736 3136491 3135856 636 prf:2518330B Corynebacterium diphtheriae 50.9 75.5 212 two-component system response chrA regulator 3237 6737 3136920 3137558 639 3238 6738 3137884 3138471 588 3239 6739 3137903 3136593 1311 prf:2518330A Corynebacterium diphtheriae 30.2 64.5 408 two-component system sensor chrS histidine kinase 3240 6740 3138630 3138481 150 gp.SCH69_22 Streptomyces coelicolor A3(2) 45.8 79.2 48 hypothetical protein SCH69.22c 3241 6741 3139455 3138634 822 gp:SCH69_20 Streptomyces coelicolor A3(2) 30.0 59.2 277 hypothetical protein SCH69.20c 3242 6742 3139651 3140952 1302 sp:SP3J_BACSU Bacillus subtilis spoIIIJ 26.0 53.6 265 stage III sporulation protein 3243 6743 3141523 3140885 639 pir:C70948 Mycobacterium tuberculosis 32.3 60.9 192 transcriptional repressor H37Rv Rv3173c 3244 6744 3141969 3141709 261 sp:TAG1_ECOLI Escherichia coli K12 MG1655 34.5 71.3 87 transglycosylase-associated protein tag1 3245 6745 3143356 3142454 903 sp:YW12_MYCTU Mycobacterium tuberculosis 41.2 69.6 296 hypothetical protein H37Rv Rv2005c 3246 6746 3144482 3143496 987 sp:YHBW_ECOLI Escherichia coli K12 MG1655 38.5 73.9 314 hypothetical protein yhbW 3247 6747 3144661 3145626 966 sp:YBC5_CHLVI Chlorobium vibrioforme ybc5 28.4 51.2 334 RNA pseudouridylate synthase 3248 6748 3146569 3146841 273 GSP:Y35814 Chlamydia pneumoniae 61.0 66.0 84 hypothetical protein 3249 6749 3147090 3147230 141 PIR:F81737 Chlamydia muridarum Nigg 71.0 75.0 42 hypothetical protein TC0129 3250 6750 3151575 3151369 207 3251 6751 3152204 3151842 363 sp:GLCC_ECOLI Escherichia coli K12 MG1655 30.3 56.0 109 bacterial regulatory protein, gntR glcC family or glc operon transcriptional activator 3252 6752 3152413 3153828 1416 gp:SC4G6_31 Streptomyces coelicolor 26.0 48.2 488 hypothetical protein SC4G6.31c 3253 6753 3154766 3153894 873 sp:35KD_MYCTU Mycobacterium tuberculosis 48.3 78.7 267 hypothetical protein H37Rv Rv2744c 3254 6754 3154817 3154969 153 3255 6755 3156697 3155246 1452 3256 6756 3157373 3156306 1068 3257 6757 3157471 3157223 249 3258 6758 3157787 3157479 309 3259 6759 3158124 3158834 711 gp:SCD35_11 Streptomyces coelicolor A3(2) 32.3 58.1 217 methyltransferase SCD35.11c 3260 6760 3159800 3159081 720 sp:NO21_SOYBN soybean NO21 26.1 55.2 241 nodulin 21-related protein 3261 6761 3160216 3160419 204 3262 6762 3160688 3161065 378 3263 6763 3160816 3161001 186 3264 6764 3160938 3160723 216 sp:TNP5_PSEAE Pseudomonas aeruginosa TNP5 48.2 92.9 56 transposon tn501 resolvase 3265 6765 3161219 3161701 483 3266 6766 3161407 3161087 321 sp:FER_SACER Saccharopolyspora erythraea fer 90.3 98.4 62 ferredoxin precursor 3267 6767 3162014 3161682 333 gp:SCD31_14 Streptomyces coelicolor A3(2) 47.3 85.5 55 hypothetical protein 3268 6768 3162694 3162804 111 GPU:AF164956_8 Corynebacterium glutamicum 81.0 84.0 27 transposase Tnp1673 3269 6769 3162710 3162871 162 GPU:AF164956_23 Corynebacterium glutamicum 84.0 90.0 46 transposase protein fragment TnpNC 3270 6770 3162852 3163889 1038 3271 6771 3162983 3162858 126 sp:G3P_PYRWO Pyrococcus woesei gap 63.2 84.2 38 glyceraldehyde-3-phosphate dehydrogenase(pseudogene) 3272 6772 3163733 3163074 660 pir:S77018 Synechocystis sp. PCC6803 32.2 59.4 180 lipoprotein sll0788 3273 6773 3166005 3163789 2217 pir:H69268 Archaeoglobus fulgidus AF0152 45.8 73.4 717 copper/potassium-transporting ATPase B or cation transporting ATPase (E1-E2 family) 3274 6774 3166437 3166267 171 3275 6775 3166978 3167169 192 3276 6776 3167646 3166450 1197 sp:BAES_ECOLI Escherichia coli K12 baeS 37.5 71.4 301 two component system sensor histidine kinase 3277 6777 3167739 3168566 828 3278 6778 3168401 3167646 756 sp:PHOP_BACSU Bacillus subtilis phoP 43.4 72.1 233 two-component response regulator or alkaline phosphatase synthesis transcriptional regulatory protein 3279 6779 3168669 3169340 672 3280 6780 3169414 3170892 1479 sp:COPA_PSESM Pseudomonas syringae pv. 26.7 47.9 630 laccase or copper resistance protein tomato copA precursor A 3281 6781 3171254 3171616 363 sp:TLPA_BRAJA Bradyrhizobium japonicum tlpA 31.7 63.4 101 thiol:disulfide interchange protein (cytochrome c biogenesis protein) 3282 6782 3172536 3171619 918 sp:QOR_MOUSE Mus musculus qor 31.4 60.9 322 quinone oxidoreductase (NADPH:quinone reductase)(seta- crystallin) 3283 6783 3172995 3173465 471 3284 6784 3173624 3173857 234 sp:ATZN_SYNY3 Synechocystis sp. PCC6803 37.2 66.7 78 zinc-transporting ATPase (Zn(II)- atzN translocating p-type ATPase 3285 6785 3174066 3174380 315 3286 6786 3174990 3174784 207 3287 6787 3175027 3176901 1875 sp: ATZN_ECOLI Escherichia coli K12 MG1655 39.8 68.5 606 zinc-transporting ATPase (Zn(II)- atzN translocating p-type ATPase 3288 6788 3175643 3175254 390 PIR:E72491 Aeropyrum pernix K1 APE2572 45.0 54.0 72 hypothetical protein 3289 6789 3177174 3177482 309 3290 6790 3177304 3177089 216 GPU:AF164956_8 Corynebacterium glutamicum 58.0 73.0 73 transposase Tnp1673 3291 6791 3177565 3177308 258 GPU:AF164956_8 Corynebacterium glutamicum 75.0 77.0 70 transposase Tnp1673 3292 6792 3177683 3177525 159 gp:AF121000_8 Corynebacterium glutamicum 92.5 96.2 53 transposase (IS1628) 22243 R-plasmid pAG1 tnpB 3293 6793 3178558 3178112 447 sp:THI2_ECOLI Escherichia coli K12 thi2 39.0 74.0 100 thioredoxin 3294 6794 3178609 3178872 264 3295 6795 3179049 3180392 1344 sp:PCAK_PSEPU Pseudomonas putida pcaK 27.1 60.1 421 transmembrane transport protein or 4-hydroxybenzoate transporter 3296 6796 3181104 3180946 159 3297 6797 3181126 3180551 576 sp:YQJI_ECOLI Escherichia coli K12 yqjl 35.1 62.5 208 hypothetical protein 3298 6798 3182866 3181337 1530 sp:DNAB_ECOLI Escherichia coli K12 dnaB 37.7 73.1 461 replicative DNA helicase 3299 6799 3183469 3183984 516 3300 6800 3183927 3183478 450 sp:RL9_ECOLI Escherichia coli K12 RL9 42.2 71.4 154 50S ribosomal protein L9 3301 6801 3184661 3183987 675 sp:SSB_ECOLI Escherichia coli K12 ssb 30.6 51.5 229 single-strand DNA binding protein 3302 6802 3184985 3184701 285 sp:RS6_ECOLI Escherichia coli K12 RS6 28.3 78.3 92 30S ribosomal protein S6 3303 6803 3185536 3185348 189 3304 6804 3186993 3185536 1458 gp:AF187306_1 Mycobacterium smegmatis 41.5 68.3 480 hypothetical protein mc(2)155 3305 6805 3187912 3188793 882 3306 6806 3189201 3187042 2160 sp:PBPA_BACSU Bacillus subtilis ponA 29.1 60.1 647 penicillin-binding protein 3307 6807 3189652 3189296 357 sp:Y0HC_MYCTU Mycobacterium tuberculosis 41.1 72.0 107 hypothetical protein H37Rv Rv0049 3308 6808 3189877 3190347 471 pir:B70912 Mycobacterium tuberculosis 35.1 65.0 137 bacterial regulatory protein, marR H37Rv Rv0042c family 3309 6809 3190378 3191319 942 sp:Y0FF_MYCTU Mycobacterium tuberculosis 29.7 61.8 296 hypothetical protein H37Rv Rv2319c yofF 3310 6810 3191354 3191848 495 3311 6811 3192242 3191922 321 sp:YHGC_BACSU Bacillus subtilis yhgC 32.4 70.4 71 hypothetical protein 3312 6812 3193201 3192266 936 sp:YCEA_ECOLI Escherichia coli K12 yceA 30.2 63.8 298 hypothetical protein 3313 6813 3194514 3193252 1263 sp:YBJZ_ECOLI Escherichia coli K12 ybjZ 31.2 64.0 433 ABC transporter ATP-binding protein 3314 6814 3195203 3194514 690 sp:YBJZ_ECOLI Escherichia coli K12 MG1655 48.9 80.1 221 ABC transporter ATP-binding protein ybjZ 3315 6815 3197186 3195210 1977 pir:E81408 Campylobacter jejuni Cj0606 18.0 42.0 237 hypothetical protein 3316 6816 3197412 3198500 1089 pir:F70912 Mycobacterium tuberculosis 77.8 90.0 360 hypothetical protein H37Rv Rv0046c 3317 6817 3199187 3198582 606 3318 6818 3200686 3199202 1485 3319 6819 3201754 3201260 495 sp:DPS_ECOLI Escherichia coli K12 dps 37.7 64.9 154 DNA protection during starvation protein 3320 6820 3201900 3202712 813 sp:FPG_ECOLI Escherichia coli K12 mutM or 28.4 55.6 268 formamidopyrimidine-DNA fpg glycosylase 3321 6821 3202952 3204100 1149 sp:RTCB_ECOLI Escherichia coli K12 rtcB 47.5 66.6 404 hypothetical protein 3322 6822 3204067 3202979 1089 3323 6823 3204156 3204728 573 3324 6824 3205204 3204731 474 sp:MGMT_HUMAN Homo sapiens mgmT 38.0 63.3 166 methylated-DNA-protein-cysteine S-methyltransferase 3325 6825 3206232 3205222 1011 sp:QOR_CAVPO Cavia porcellus (Guinea pig) qor 33.3 63.6 231 zinc-binding dehydrogenase or quinone oxidoreductase (NADPH:quinone reductase) or alginate lyase 3326 6826 3206646 3206756 111 3327 6827 3206849 3208024 1176 sp:YDEA_ECOLI Mycobacterium tuberculosis 26.4 66.3 398 membrane transport protein H37Rv Rv0191 ydeA 3328 6828 3208279 3209454 1176 gp:AF234535_1 Corynebacterium melassecola 99.7 99.5 392 malate oxidoreductase [NAD](malic (Corynebacterium glutamicum) enzyme) ATCC 17965 malE 3329 6829 3211186 3209705 1482 sp:GNTK_BACSU Bacillus subtilis gntK 24.5 53.7 486 gluconokinase or gluconate kinase 3330 6830 3211836 3211246 591 sp:VANZ_ENTFC Enterococcus faecium vanZ 27.8 60.4 169 teicoplanin resistance protein 3331 6831 3212428 3211904 525 sp:VANZ_ENTFC Enterococcus faecium vanZ 27.0 159.0 159 teicoplanin resistance protein 3332 6832 3212588 3213931 1344 sp:MERA_STAAU Staphylococcus aureus merA 29.9 65.6 448 mercury(II) reductase 3333 6833 3215163 3213934 1230 sp:DADA_ECOLI Escherichia coli K12 dadA 27.3 54.5 444 D-amino acid dehydrogenase small subunit 3334 6834 3216759 3215257 1503 3335 6835 3217215 3216886 330 3336 6836 3217777 3217457 321 3337 6837 3217993 3218601 609 sp:NOX_THETH Thermus thermophilus nox 25.8 55.2 194 NAD(P)H nitroreductase 3338 6838 3218777 3219700 924 3339 6839 3221044 3222495 1452 3340 6840 3222633 3219778 2856 sp:SYL_BACSU Bacillus subtilis syl 47.7 68.1 943 leucyl-tRNA synthetase 3341 6841 3222722 3223150 429 sp:YBAN_ECOLI Escherichia coli K12 40.4 40.4 104 hypothetical membrane protein 3342 6842 3223445 3223089 357 sp:VAPI_BACNO Dichelobacter nodosus vapl 55.8 81.4 86 virulence-associated protein 3343 6843 3224601 3225374 774 3344 6844 3224714 3223992 723 gp:SCC54_19 Streptomyces coelicolor 31.6 53.8 247 hypothetical protein SCC54.19 3345 6845 3225554 3224718 837 sp:HPCE_ECOLI Escherichia coli K12 hpcE 28.5 50.3 298 bifunctional protein (homoprotocatechuate catabolism bifunctional isomerase/decarboxylase) (2- hydroxyhepta-2,4-diene-1,7-dioate isomerase and 5-carboxymethyl-2- oxo-hex-3-ene-1,7dioate decarboxylase) 3346 6846 3226687 3225563 1125 gp:AF173167_1 Pseudomonas alcaligenes xlnE 34.2 64.3 339 gentisate 1,2-dioxygenase or 1- hydroxy-2-naphthoate dioxygenase 3347 6847 3227689 3226910 780 sp:KDGR_ERWCH Pectobacterium chrysanthemi 25.3 60.7 229 bacterial regulatory protein, lacl kdgR family or pectin degradation repressor protein 3348 6848 3227724 3229079 1356 sp:PCAK_PSEPU Pseudomonas putida pcaK 27.5 60.8 454 transmembrane transport protein or 4-hydroxybenzoate transporter 3349 6849 3229119 3230444 1326 prf:1706191A Pseudomonas putida 28.2 49.4 476 salicylate hydroxylase 3350 6850 3232304 3231054 1251 sp:EAT2_HUMAN Homo sapiens eat2 25.4 54.4 507 proton/glutamate symporter or excitatory amino acid transporter2 3351 6851 3232596 3233105 510 pir:JC2326 Corynebacterium glutamicum 99.4 99.4 170 tryptophan-specific permease AS019 ORF1 3352 6852 3233403 3234956 1554 sp:TRPE_BRELA Brevibacterium lactofermentum 99.2 99.8 515 anthranilate synthase component I trpE 3353 6853 3233420 3233250 171 3354 6854 3234956 3235579 624 TRPG_BRELA Brevibacterium lactofermentum 99.0 100.0 208 anthranilate synthase component II trpG 3355 6855 3235602 3236645 1044 sp:TRPD_CORGL Corynebacterium glutamicum 99.4 99.4 348 anthranilate ATCC 21850 trpD phosphoribosyltransferase 3356 6856 3236641 3238062 1422 sp:TRPC_BRELA Brevibacterium lactofermentum 97.3 98.3 474 indole-3-glycerol phosphate trpC synthase (IGPS) and N-(5′- phosphoribosyl) anthranilate isomerase(PRAI) 3357 6857 3237213 3236518 696 3358 6858 3238082 3239332 1251 sp:TRPB_BRELA Brevibacterium lactofermentum 97.6 97.9 417 tryptophan synthase beta chain trpB 3359 6859 3239332 3240171 840 sp:TRPA_BRELA Brevibacterium lactofermentum 95.4 96.5 283 tryptophan synthase alpha chain trpA 3360 6860 3241851 3240313 1539 gp:SCJ21_17 Streptomyces coelicolor A3(2) 66.6 86.8 521 hypothetical membrane protein SCJ21.17c 3361 6861 3242688 3241879 810 sp:PTXA_ECOLI Escherichia coli K12 ptxA 30.3 71.7 152 PTS system, IIA component or unknown pentitol phosphotransferase enzyme II, A component 3362 6862 3242854 3243759 906 sp:NOSF_PSEST Pseudomonas stutzeri 32.5 63.6 305 ABC transporter ATP-binding protein 3363 6863 3243759 3245342 1584 gp:SCH10_12 Streptomyces coelicolor A3(2) 25.2 57.2 547 ABC transporter SCH10.12 3364 6864 3245317 3245766 450 sp:UCRI_CHLLT Chlorobium limicola petC 32.5 63.6 305 cytchrome b6-F complex iron-sulfur subunit (Rieske iron-sulfur protein) 3365 6865 3246931 3245822 1110 sp:NADO_THEBR Thermoanaerobacter brockii 33.3 64.3 336 NADH oxidase or NADH-dependent nadO flavin oxidoreductase 3366 6866 3247234 3248205 972 sp:YFEH_ECOLI Escherichia coli K12 yfeH 43.6 74.7 328 hypothetical membrane protein 3367 6867 3248392 3249165 774 gp:SCI11_36 Streptomyces coelicolor A3(2) 34.0 54.6 262 hypothetical protein SCI11.36c 3368 6868 3249534 3249187 348 pir:A29606 Streptomyces coelicolor Plasmid 45.1 79.4 102 bacterial regulatory protein, arsR SCP1 mmr family or methylenomycin A resistance protein 3369 6869 3249651 3250742 1092 sp:NADO_THEBR Thermoanaerobacter brockii 33.4 64.3 347 NADH oxidase or NADH-dependent nadO flavin oxidoreductase 3370 6870 3250758 3251405 648 sp:YMY0_YEAST Saccharomyces cerevisiae 31.4 69.5 226 hypothetical protein ymyO 3371 6871 3251618 3251466 153 3372 6872 3251934 3251743 192 3373 6873 3252300 3252133 168 3374 6874 3252636 3252316 321 3375 6875 3252728 3253480 753 sp:BUDC_KLETE Klebsiella terrigena budC 26.9 52.9 238 acetoin(diacetyl) reductase (acetoin dehydrogenase) 3376 6876 3253560 3253739 180 sp:YY34_MYCTU Mycobacterium tuberculosis 53.5 84.5 58 hypothetical protein H37Rv Rv2094c 3377 6877 3255182 3253824 1359 sp:DTPT_LACLA Lactococcus lactis subsp. lactis 34.5 71.6 469 di-/tripeptide transpoter dtpT 3378 6878 3255549 3255719 171 3379 6879 3256298 3255744 555 sp:ACRR_ECOLI Escherichia coli K12 acrR 26.1 50.5 188 bacterial regulatory protein, tetR family 3380 6880 3257373 3256471 903 sp:CATA_ACICA Acinetobacter calcoaceticus 31.7 62.2 246 hydroxyquinol 1,2-dioxygenase catA 3381 6881 3258491 3257403 1089 sp:TCBF_PSESQ Pseudomonas sp. P51 43.0 75.5 351 maleylacetate reductase 3382 6882 3260084 3258561 1524 sp:XYLE_ECOLI Escherichia coli K12 xylE 31.4 58.3 513 sugar transporter or D-xylose-proton symporter (D-xylose transporter) 3383 6883 3261129 3261989 861 sp:ICLR_SALTY Salmonella typhimurium iclR 25.7 60.7 280 bacterial transcriptional regulator or acetate operon repressor 3384 6884 3262145 3263221 1077 sp:YDGJ_ECOLI Escherichia coli K12 ydgJ 27.2 55.7 357 oxidoreductase 3385 6885 3263237 3264115 879 gsp:W61761 Listeria innocua strain 4450 25.9 58.2 270 diagnostic fragment protein sequence 3386 6886 3264142 3265146 1005 sp:MI2D_BACSU Sinorhizobium meliloti idhA 26.5 59.6 332 myo-inositol 2-dehydrogenase 3387 6887 3265184 3266266 1083 sp:STRI_STRGR Streptomyces griseus strl 34.1 62.4 343 dehydrogenase or myo-inositol 2- dehydrogenase or streptomycin biosynthesis protein 3388 6888 3267062 3271093 4032 pir:C70044 Bacillus subtilis yvnB 33.3 62.7 1242 phosphoesterase 3389 6889 3268557 3267913 645 3390 6890 3269235 3268618 618 3391 6891 3271392 3272477 1086 3392 6892 3275231 3274488 744 SP:UNC1_CAEEL Caenorhabditis elegans unc1 28.6 57.3 206 stomatin 3393 6893 3276570 3275602 969 3394 6894 3281599 3276671 4929 gp:MBO18605_3 Mycobacterium bovis BCG 58.4 80.2 1660 DEAD box RNA helicase family RvD1-Rv2024c 3395 6895 3282172 3281666 507 prf:2323363AAM Mycobacterium leprae u2266k 34.8 61.0 141 hypothetical membrane protein 3396 6896 3282742 3283101 360 3397 6897 3282946 3282347 600 sp:THID_BACSU Bacillus subtilis thiD 50.4 76.8 125 phosphomethylpyrimidine kinase 3398 6898 3283141 3283383 243 pir:F70041 Bacillus subtillis yvgY 46.3 70.1 67 mercuric ion-binding protein or heavy-metal-associated domain containing protein 3399 6899 3284309 3283473 837 prf:2501295A Corynebacterium glutamicum 29.9 62.3 297 ectoine/proline uptake protein proP 3400 6900 3285355 3284399 957 sp:FECB_ECOLI Escherichia coli K12 fecB 29.4 60.6 279 iron(III) dicitrate-binding periplasmic protein precursor or iron(III) dicitrate transport system permease protein 3401 6901 3285455 3286576 1122 sp:MRF1_SCHPO Schizosaccharomyces pombe 27.2 58.0 324 mitochondrial respiratory function mrf1 protein or zinc-binding dehydrogenase or NADPH quinone oxidoreductase 3402 6902 3286622 3287005 384 3403 6903 3287297 3287079 219 3404 6904 3288190 3287393 798 sp:THID_BACSU Bacillus subtilis thiD 46.2 75.5 249 phosphomethylpyrimidine kinase 3405 6905 3288265 3288609 345 3406 6906 3288685 3288885 201 pir:F70041 Bacillus subtilis yvgV 41.8 70.1 67 mercuric ion-binding protein or heavy-metal-associated domain containing protein 3407 6907 3289315 3288971 345 sp:AZLD_BACSU Bacillus subtilis azlD 36.3 65.7 102 branched-chain amino acid transport 3408 6908 3290021 3289311 711 sp:AZLC_BACSU Bacillus subtilis azlD 32.1 67.0 212 branched-chain amino acid transport 3409 6909 3290591 3290025 567 sp:YQGE_ECOLI Escherichia coli K12 yqgE 23.7 56.2 169 hypothetical protein 3410 6910 3291942 3290623 1320 sp:CCA_ECOLI Escherichia coli K12 cca 26.8 51.8 471 tRNA nucleotidyltransferase 3411 6911 3292532 3293497 966 pir:E70600 Mycobacterium tuberculosis 43.6 69.2 234 mutator mutT protein H37Rv Rv3908 3412 6912 3292882 3292610 273 3413 6913 3293497 3296007 2511 pir:F70600 Mycobacterium tuberculosis 25.8 54.3 858 hypothetical membrane protein H37Rv Rv3909 3414 6914 3296156 3299404 3249 pir:G70600 Mycobacterium tuberculosis 35.7 60.1 1201 hypothetical membrane protein H37Rv Rv3910 3415 6915 3297706 3298428 723 3416 6916 3299661 3300263 603 sp:RPSH_PSEAE Pseudomonas aeruginosa algU 30.2 60.9 189 RNA polymerase sigma-H factor or sigma-70 factor (ECF subfamily) 3417 6917 3300371 3301321 951 sp:TRXB_STRCL Streptomyces clavuligerus trxB 60.4 82.5 308 thioredoxin reductase 3418 6918 3301303 3300119 1185 3419 6919 3301358 3301729 372 sp:THI2_CHLRE Chlamydomonas reinhardtii thi2 42.0 76.5 119 thioredoxin ch2, M-type 3420 6920 3301755 3302996 1242 sp:CWLB_BACSU Bacillus subtilis cwlB 51.0 75.4 196 N-acetylmuramoyl-L-alanine amidase 3421 6921 3302765 3301989 777 3422 6922 3303435 3304475 1041 3423 6923 3303616 3302999 618 pir:D70851 Mycobacterium tuberculosis 34.4 58.5 212 hypothetical protein H37Rv Rv3916c 3424 6924 3304787 3303636 1152 sp:YGI2_PSEPU Pseudomonas putida ygi2 37.6 60.5 367 hypothetical protein 3425 6925 3305671 3304835 837 sp:YGI1_PSEPU Mycobacterium tuberculosis 65.0 78.0 272 partitioning or sporulation protein H37Rv parB 3426 6926 3306532 3305864 669 sp:GIDB_ECOLI Escherichia coli K12 gidB 36.0 64.7 153 glucose inhibited division protein B 3427 6927 3307632 3306682 951 pir:A70852 Mycobacterium tuberculosis 44.7 75.4 313 hypothetical membrane protein H37Rv Rv3921c 3428 6928 3308369 3307971 399 sp:RNPA_BACSU Bacillus subtilis rnpA 26.8 59.4 123 ribonuclease P protein component 3429 6929 3308747 3308412 336 gp:MAU19185_1 Mycobacterium avium rpmH 83.0 93.6 47 50S ribosomal protein L34 3430 6930 3309028 3309321 294 3431 6931 3309043 3308822 222 3432 6932 147980 147573 408 gp:AF116184_1 Corynebacterium glutamicum 100.0 100.0 136 L-aspartate-alpha-decarboxylase panD precursor 3433 6933 268001 266154 1848 sp:LEU1_CORGL Corynebacterium glutamicum 100.0 100.0 616 2-isopropylmalate synthase ATCC 13032 leuA 3434 6934 269068 268814 255 sp:YLEU_CORGL Corynebacterium glutamicum 100.0 100.0 85 hypothetical protein (Brevibacterium flavum) ATCC 13032 orfX 3435 6935 270660 271691 1032 sp:DHAS_CORGL Corynebacterium glutamicum 100.0 100.0 344 aspartate-semialdehyde asd dehydrogenase 3436 6936 446075 446521 447 gp:AF124518_1 Corynebacterium glutamicum 100.0 100.0 149 3-dehydroquinase ASO19 aroD 3437 6937 526376 527563 1188 sp:EFTU_CORGL Corynebacterium glutamicum 100.0 100.0 396 elongation factorTu ATCC 13059 tuf 3438 6938 569452 570771 1320 sp:SECY_CORGL Corynebacterium glutamicum 100.0 100.0 440 preprotein translocase secY subuit (Brevibacterium flavum) MJ233 secY 3439 6939 680044 677831 2214 sp:IDH_CORGL Corynebacteriurn glutamicum 100.0 100.0 738 isocitrate dehydrogenase ATCC 13032 icd (oxalosuccinatedecarboxylase) 3440 6940 720352 718580 1773 prf:2223173A Corynebacterium glutamicum 100.0 100.0 591 acyl-CoA carboxylase or biotin- ATCC 13032 accBC binding protein 3441 6941 877838 879148 1311 sp:CISY_CORGL Corynebacterium glutamicum 100.0 100.0 437 citrate synthase ATCC 13032 gltA 3442 6942 879276 879629 354 sp:FKBP_CORGL Corynebacterium glutamicum 100.0 100.0 118 putative binding protein or peptidyl- ATCC 13032 fkbA prolyl cis-trans isomerase 3443 6943 944996 946780 1785 sp:BETP_CORGL Corynebacterium glutamicum 100.0 100.0 595 glycine betaine transporter ATCC 13032 betP 3444 6944 1030283 1029006 1278 sp:YLI2_CORGL Corynebacterium glutamicum 100.0 100.0 426 hypothetical membrane protein ATCC 13032 orf2 3445 6945 1031871 1030369 1503 sp:LYSI_CORGL Corynebacterium glutamicum 100.0 100.0 501 L-lysine permease ATCC 13032 lysI 3446 6946 1154683 1153295 1389 sp:AROP_CORGL Corynebacterium glutamicum 100.0 100.0 463 aromatic amino acid permease ATCC 13032 aroP 3447 6947 1155676 1154729 948 pir:S52753 Corynebacterium glutamicum 100.0 100.0 316 hypothetical protein ATCC 13032 orf3 3448 6948 1155731 1156837 1107 prf:2106301A Corynebacterium glutamicum 100.0 100.0 369 succinyl diaminopimelate ATCC 13032 dapE desuccinylase 3449 6949 1219602 1218031 1572 gp:CGPUTP_1 Corynebacterium glutamicum 100.0 100.0 524 proline transport system ATCC 13032 putP 3450 6950 1238274 1239923 1650 sp:SYR_CORGL Corynebacterium glutamicum 100.0 100.0 550 arginyl-tRNA synthetase AS019 ATCC 13059 argS 3451 6951 1239929 1241263 1335 sp:DCDA_CORGL Corynebacterium glutamicum 100.0 100.0 445 diaminopimelate (DAP) AS019 ATCC 13059 lysA decarboxylase (meso- diaminopimelate decarboxylase) 3452 6952 1242507 1243841 1335 sp:DHOM_CORGL Corynebacterium glutamicum 100.0 100.0 445 homoserine dehydrogenase AS019 ATCC 13059 hom 3453 6953 1243855 1244781 927 sp:KHSE_CORGL Corynebacterium glutamicum 100.0 100.0 309 homoserine kinase AS019 ATCC 13059 thrB 3454 6954 1327617 1328243 627 gsp:W37716 Corynebacterium glutamicum 100.0 100.0 216 ion channel subunit R127 orf3 3455 6955 1328953 1328246 708 sp:LYSE_CORGL Corynebacterium glutamicum 100.0 100.0 236 lysine exporter protein R127 lysE 3456 6956 1329015 1329884 870 sp:LYSG_CORGL Corynebacterium glutamicum 100.0 100.0 290 lysine export regulator protein R127 lysG 3457 6957 1338131 1340008 1878 sp:ILVB_CORGL Corynebacterium glutamicum 100.0 100.0 626 acetohydroxy acid synthase, large ATCC 13032 ilvB subunit 3458 6958 1340025 1340540 516 pir:B48648 Corynebacterium glutamicum 100.0 100.0 172 acetohydroxy acid synthase, small ATCC 13032 ilvN subunit 3459 6959 1340724 1341737 1014 pir:C48648 Corynebacterium glutamicum 100.0 100.0 338 acetohydroxy acid isomeroreductase ATCC 13032 ilvC 3460 6960 1353489 1354508 1020 sp:LEU3_CORGL Corynebacterium glutamicum 100.0 100.0 340 3-isopropylmalate dehydrogenase ATCC 13032 leuB 3461 6961 1423217 1425265 2049 prf:2014259A Corynebacterium glutamicum 100.0 100.0 683 PTS system, phosphoenolpyruvate KCTC1445 ptsM sugar phosphotransferase (mannose and glucose transport) 3462 6962 1466491 1467372 882 sp:ARGB_CORGL Corynebacterium glutamicum 100.0 100.0 294 acetylglutamate kinase ATCC 13032 argB 3463 6963 1468565 1469521 957 sp:OTCA_CORGL Corynebacterium glutamicum 100.0 100.0 319 ornithine carbamoyltransferase ATCC 13032 argF 3464 6964 1469528 1470040 513 gp.AF041436_1 Corynebacterium glutamicum 100.0 100.0 171 arginine repressor ASO19 argR 3465 6965 1544554 1543154 1401 gp:CGL238250_1 Corynebacterium glutamicum 100.0 100.0 467 NADH dehydrogenase ATCC 13032 ndh 3466 6966 1586725 1586465 261 gp:AF086704_1 Corynebacterium glutamicum 100.0 100.0 87 phosphoribosyl-ATP- ASO19 hisE pyrophosphohydrolase 3467 6967 1675208 1674123 1086 gp:CGL007732_4 Corynebacterium glutamicum 100.0 100.0 362 ornithine-cyclodecarboxylase ATCC 13032 ocd 3468 6968 1676623 1675268 1356 gp:CGL007732_3 Corynebacterium glutamicum 100.0 100.0 452 ammonium uptake protein, high ATCC 13032 amt affinity 3469 6969 1677279 1677049 231 gp:CGL007732_2 Corynebacterium glutamicum 100.0 100.0 77 protein-export membrane protein ATCC 13032 secG secG 3470 6970 1680143 1677387 2757 prf:1509267A Corynebacterium glutamicum 100.0 100.0 919 phosphoenolpyruvate carboxylase ATCC 13032 ppc 3471 6971 1720898 1719669 1230 gp:AF124600_1 Corynebacterium glutamicum 100.0 100.0 410 chorismate synthase (5- AS019 aroC enolpyruvylshikimate-3-phosphate phospholyase) 3472 6972 1880490 1882385 1896 pir:B55225 Corynebacterium glutamicum 100.0 100.0 632 restriction endonuclease ATCC 13032 cgIIIR 3473 6973 2020854 2021846 993 prf:2204286D Corynebacterium glutamicum 100.0 100.0 331 sigma factor or RNA polymerase ATCC 13869 sigB transcription factor 3474 6974 2060620 2061504 885 sp:GLUB_CORGL Corynebacterium glutamicum 100.0 100.0 295 glutamate-binding protein ATCC 13032 gluB 3475 6975 2065116 2063989 1128 sp:RECA_CORGL Corynebacterium glutamicum 100.0 100.0 376 recA protein AS019 recA 3476 6976 2080183 2079281 903 sp:DAPA_BRELA Corynebacterium glutamicum 100.0 100.0 301 dihydrodipicolinate synthase (Brevibacterium lactofermentum) ATCC 13869 dapA 3477 6977 2081934 2081191 744 sp:DAPB_CORGL Corynebacterium glutamicum 100.0 100.0 248 dihydrodipicolinate reductase (Brevibacterium lactofermentum) ATCC 13869 dapB 3478 6978 2115363 2113864 1500 gp:CGA224946_1 Corynebacterium glutamicum 100.0 100.0 500 L-malate dehydrogenase (acceptor) R127 mqo 3479 6979 2171741 2169666 2076 gp:CAJ10319_4 Corynebacterium glutamicum 100.0 100.0 692 uridilylyltransferase, uridilylyl- ATCC 13032 glnD removing enzyme 3480 6980 2172086 2171751 336 gp:CAJ10319_3 Corynebacterium glutamicum 100.0 100.0 112 nitrogen regulatory protein P-II ATCC 13032 glnB 3481 6981 2173467 2172154 1314 gp:CAJ10319_2 Corynebacterium glutamicum 100.0 100.0 438 ammonium transporter ATCC 13032 amtP 3482 6982 2196082 2194742 1341 pir:S32227 Corynebacterium glutamicum 100.0 100.0 447 glutamate dehydrogenase (NADP+) ATCC 17965 gdhA 3483 6983 2207092 2205668 1425 sp:KPYK_CORGL Corynebacterium glutamicum 100.0 100.0 475 pyruvate kinase ASO19 pyk 3484 6984 2317550 2316582 969 gp:AF096280_1 Corynebacterium glutamicum 100.0 100.0 323 glucokinase ATCC 13032 glk 3485 6985 2348829 2350259 1431 prf:2322244A Corynebacterium glutamicum 100.0 100.0 477 glutamine synthetase ATCC 13032 glnA 3486 6986 2355042 2353600 1443 sp:THRC_CORGL Corynebacterium glutamicum 100.0 100.0 481 threonine synthase thrC 3487 6987 2450172 2448328 1845 prf:2501295B Corynebacterium glutamicum 100.0 100.0 615 ectoine/proline/glycine betaine ATCC 13032 ectP carrier 3488 6988 2470141 2467925 2217 pir:I40715 Corynebacterium glutamicum 100.0 100.0 739 malate synthase ATCC 13032 aceB 3489 6989 2470740 2472035 1296 pir:I40713 Corynebacterium glutamicum 100.0 100.0 432 isocitrate lyase ATCC 13032 aceA 3490 6990 2497776 2496670 1107 sp:PROB_CORGL Corynebacterium glutamicum 100.0 100.0 369 glutamate 5-kinase ATCC 17965 proB 3491 6991 2591469 2590312 1158 gp:AF126953_1 Corynebacterium glutamicum 100.0 100.0 386 cystathionine gamma-synthase ASO19 metB 3492 6992 2680127 2679684 444 gp:AF112535_2 Corynebacterium glutamicum 100.0 100.0 148 ribonucleotide reductase ATCC 13032 nrdl 3493 6993 2680649 2680419 231 gp:AF112535_1 Corynebacterium glutamicum 100.0 100.0 77 glutaredoxin ATCC 13032 nrdH 3494 6994 2787715 2786756 960 sp.DDH_CORGL Corynebacterium glutamicum 100.0 100.0 320 meso-diaminopimelate D- KY10755 ddh dehydrogenase 3495 6995 2888078 2887944 135 gp:CGL238703_1 Corynebacterium glutamicum 100.0 100.0 45 porin or cell wall channel forming MH20-22B porA protein 3496 6996 2936505 2935315 1191 sp:ACKA_CORGL Corynebacterium glutamicum 100.0 100.0 397 acetate kinase ATCC 13032 ackA 3497 6997 2937494 2936508 987 prf:2516394A Corynebacterium glutamicum 100.0 100.0 329 phosphate acetyltransferase ATCC 13032 pta 3498 6998 2961342 2962718 1377 prf:2309322A Corynebacterium glutamicum 100.0 100.0 459 multidrug resistance protein or ATCC 13032 cmr macrolide-efflux pump or drug:proton antiporter 3499 6999 2966161 2963606 2556 sp:CLPB_CORGL Corynebacterium glutamicum 100.0 100.0 852 ATP-dependent protease regulatory ATCC 13032 clpB subunit 3500 7000 3099522 3098578 945 prf:1210266A Corynebacterium glutamicum 100.0 100.0 315 prephenate dehydratase pheA 3501 7001 3274074 3272563 1512 prf:2501295A Corynebacterium glutamicum 100.0 100.0 504 ectoine/proline uptake protein ATCC 13032 proP
EXAMPLE 2[0597] Determination of effective mutation site
[0598] (1) Identification of mutation site based on the comparison of the gene nucleotide sequence of lysine-producing B-6 strain with that of wild type strain ATCC 13032
[0599] Corynebacterium glutamicum B-6, which is resistant to S-(2-aminoethyl)cysteine (AEC), rifampicin, streptomycin and 6-azauracil, is a lysine-producing mutant having been mutated and bred by subjecting the wild type ATCC 13032 strain to multiple rounds of random mutagenesis with a mutagen, N-methyl-N′-nitro-N-nitrosoguanidine (NTG) and screening (Appl. Microbiol. Biotechnol., 32:269-273 (1989)). First, the nucleotide sequences of genes derived from the B-6 strain and considered to relate to the lysine production were determined by a method similar to the above. The genes relating to the lysine production include lyse and lysG which are lysine-excreting genes; ddh, dapA, hom and lysC (encoding diaminopimelate dehydrogenase, dihydropicolinate synthase, homoserine dehydrogenase and aspartokinase, respectively) which are lysine-biosynthesis genes; and pyc and zwf (encoding private carboxylase and glucose-6-phosphate dehydrogenase, respectively) which are glucose-metabolizing genes. The nucleotide sequences of the genes derived from the production strain were compared with the corresponding nucleotide sequences of the ATCC 13032 strain genome represented by SEQ ID NOS:1 to 3501 and analyzed. As a result, mutation points were observed in many genes. For example, no mutation site was observed in lysE, lysG, ddh, dapA, and the like, whereas amino acid replacement mutations were found in hom, lysC, pyc, zvf, and the like. Among these mutation points, those which are considered to contribute to the production were extracted on the basis of known biochemical or genetic information. Among the mutation points thus extracted, a mutation, Val59Ala, in hor and a mutation, Pro458Ser, in pyc were evaluated whether or not the mutations were effective according to the following method.
[0600] (2) Evaluation of mutation, Val59Ala, in homn and mutation, Pro458Ser, in pyc
[0601] It is known that a mutation in hom inducing requirement or partial requirement for homoserine imparts lysine productivity to a wild type strain (Amino Acid Fermentation, ed. by Hiroshi Aida et al., Japan Scientific Societies Press). However, the relationship between the mutation, Vai59Ala, in hom and lysine production is not known. It can be examined whether or not the mutation, Val59Ala, in hom is an effective mutation by introducing the mutation to the wild type strain and examining the lysine productivity of the resulting strain. On the other hand, it can be examined whether or not the mutation, Pro458Ser, in pyc is effective by introducing this mutation into a lysine-producing strain which has a deregulated lysine-bioxynthetic pathway and is free from the pyc mutation, and comparing the lysine productivity of the resulting strain with the parent strain. As such a lysine-producing bacterium, No. 58 strain (FERM BP-7134) was selected (hereinaf ter referred to the “lysine-producing No. 58 strain” or the “No. 58 strain”). Based on the above, it was determined that the mutation, Val59Ala, in hom and the mutation, Pro458Ser, in pyc were introduced into the wild type strain of Corynebacterium glutamicum ATCC 13032 (hereinafter referred to as the “wild type ATCC 13032 strain” or the “ATCC 13032 strain”) and the lysine-producing No. 58 strain, respectively, using the gene replacement method. A plasmid vector pCES30 for the gene replacement for the introduction was constructed by the following method.
[0602] A plasmid vector pCE53 having a kanamycin-resistant gene and being capable of autonomously replicating in Coryneform bacteria (Mol. Gen. Genet., 196:175-178 (1984)) and a plasmid pMOB3 (ATCC 77282) containing a levansucrase gene (sacB) of Bacillus subtilis (Molecular Microbiology, 6:1195-1204 (1992)) were each digested with PstI. Then, after agarose gel electrophoresis, a pCE53 fragment and a 2.6 kb DNA fragment containing sacB were each extracted and purified using GENECLEAN Kit (manufactured by BIO 101). The pCE53 fragment and the 2.6 kb DNA fragment were ligated using Ligation Kit ver. 2 (manufactured by Takara Shuzo), introduced into the ATCC 13032 strain by the electroporation method (FEMS Microbiology Letters, 65:299 (1989)), and cultured on BYG agar medium (medium prepared by adding 10 g of glucose, 20 g of peptone (manufactured by Kyokuto Pharmaceutical), 5 g of yeast extract (manufactured by Difco), and 16 g of Bactoagar (manufactured by Difco) to 1 liter of water, and adjusting its pH to 7.2) containing 25 &mgr;g/ml kanamycin at 30° C. for 2 days to obtain a transformant acquiring kanamycin-resistance. As a result of digestion analysis with restriction enzymes, it was confirmed that a plasmid extracted from the resulting transformant by the alkali SDS method had a structure in which the 2.6 kb DNA fragment had been inserted into the PstI site of pCE53. This plasmid was named pCES30.
[0603] Next, two genes having a mutation point, hom and pyc, were amplified by PCR, and inserted into pCES30 according to the AT cloning method (Bio Experiment Illustrated vol. 3, published by Shujunsha). Specifically, pCES30 was digested with BamHI (manufactured by Takara Shuzo), subjected to an agarose gel electrophoresis, and extracted and purified using GENECLEAN Kit (manufactured by BIO 101). The both ends of the resulting pCES30 fragment were blunted with DNA Blunting Kit (manufactured by Takara Shuzo) according to the attached protocol. The blunt-ended pCES30 fragment was concentrated by extraction with phenol/chloroform and precipitation with ethanol, and allowed to react in the presence of Taq polymerase (manufactured by Roche Diagnostics) and dTTP at 70° C. for 2 hours so that a nucleotide, thymine (T), was added to the 3′-end to prepare a T vector of pCES30.
[0604] Separately, chromosomal DNA was prepared from the lysine-producing B-6 strain according to the method of Saito et al. (Biochem. Biophys. Acta, 72:619 (1963)). Using the chromosomal DNA as a template, PCR was carried out with Pfu turbo DNA polymelase (manufactured by stratagene). In the mutated hom gene, the DNAs having the nucleotide sequences represented by SEQ ID NOS:7002 and .7003 were used as the primer set. In the mutated pyc gene, the DNAs having the nucleotide sequences represented by SEQ ID NOS:7004 and 7005 were used as the primer set. The resulting PCR product was subjected to agarose gel electrophoresis, and extracted and purified using GENEGLEAN Kit (manufactured by BIO 101). Then, the PCR product was allowed to react in the presence of Taq polymerase (manufactured by Roche Diagnostics) and DATP at 72° C. for 10 minutes so that a nucleotide, adenine (A), was added to the 3′-end.
[0605] The above pCES30 T vector fragment and the mutated hom gene (1.7 kb) or mutated pyc gene (3.6 kb) to which the nucleotide A had been added of the PCR product were concentrated by extraction with phenol/chloroform and precipitation with ethanol, and then ligated using Ligation Kit ver. 2. The ligation products were introduced into the ATCC 13032 strain according to the electroporation method, and cultured on BYG agar medium containing 25 &mgr;g/ml kanamycin at 30° C. for 2 days to obtain kanamycin-resistant transformants. Each of the resulting transformants was cultured overnight in BYG liquid medium containing 25 &mgr;g/ml kanamycin, and a plasmid was extracted from the culturing solution medium according to the alkali SDS method. As a result of digestion analysis using restriction enzymes, it was confirmed that the plasmid had a structure in which the 1.7 kb or 3.6 kb DNA fragment had been inserted into pCES30. The plasmids thus constructed were named respectively pChom59 and pCpyc458.
[0606] The introduction of the mutations to the wild type ATCC 13032 strain and the lysine-producing No. 58 strain according to the gene replacement method was carried out according to the following method. Specifically, pChom59 and pCpyc458 were introduced to the ATCC 13032 strain and the No. 58 strain, respectively, and strains in which the plasmid is integrated into the chromosomal DNA by homologous recombination were selected using the method of Ikeda et al. (Microbiology 144:1863 (1998)). Then, the stains in which the second homologous recombination was carried out were selected by a selection method, making use of the fact that the Bacillus subtilis levansucrase encoded by pCES30 produced a suicidal substance (J. of Bacteriol., 174:5462 (1992)). Among the selected strains, strains in which the wild type hom and pyc genes possessed by the ATCC 13032 strain and the No. 58 strain were replaced with the mutated hom and pyc genes, respectively, were isolated. The method is specifically explained below.
[0607] One strain was selected from the transformants containing the plasmid, pChom59 or pCpyc458, and the selected strain was cultured in BYG medium containing 20 gg/ml kanamycin, and pCGll (Japanese Published Examined Patent Application No. 91827/94) was introduced thereinto by the electroporation method. pCGll is a plasmid vector having a spectinomycin-resistant gene and a replication origin which is the same as pCE53. After introduction of the pCGll, the strain was cultured on BYG agar medium containing 20 &mgr;g/ml kanamycin and 100 &mgr;g/ml spectinomycin at 30° C. for 2 days to obtain both the kanamycin and spectinomycin-resistant transformant. The chromosome of one strain of these transformants was examined by the Southern blotting hybridization according to the method reported by Ikeda et al. (Microbiology, 144:1863 (1998)). As a result, it was confirmed that pChom59 or pCpyc458 had been integrated into the chromosome by the homologous recombination of the Cambell type. In such a strain, the wild type and mutated hom or pyc genes are present closely on the chromosome, and the second homologous recombination is liable to arise therebetween.
[0608] Each of these transformants (having been recombined once) was spread on Suc agar medium (medium prepared by adding 100 g of sucrose, 7 g of meat extract, 10 g of peptone, 3 g of sodium chloride, 5 g of yeast extract (manufactured by Difco), and 18 g of Bactoagar (manufactured by Difco) to 1 liter of water, and adjusting its pH 7.2) and cultured at 30° C. for a day. Then the colonies thus growing were selected in each case. Since a strain in which the sacB gene is present converts sucrose into a suicide substrate, it cannot grow in this medium (J. Bacteriol., 174:5462 (1992)). On the other hand, a strain in which the sacb gene was deleted due to the second homologous recombination between the wild type and the mutated hom or pyc genes positioned closely to each other forms no suicide substrate and, therefore, can grow in this medium. In the homologous recombination, either the wild type gene or the mutated gene is deleted together with the sacB gene. When the wild type is deleted together with the sacb gene, the gene replacement into the mutated type arises.
[0609] Chromosomal DNA of each the thus obtained second recombinants was prepared by the above method of Saito et al. PCR was carried out using Pfu turbo DNA polymerase (manufactured by Stratagene) and the attached buffer. In the hom gene, DNAs having the nucleotide sequences represented by SEQ ID NOS:7002 and 7003 were used as the primer set. Also, in the pyc gene was used, DNAs having the nucleotide sequences represented by SEQ ID NOS:7004 and 7005 were used as the primer set. The nucleotide sequences of the PCR products were determined by the conventional method so that it was judged whether the hom or pyc gene of the second recombinant was a wild type or a mutant. As a result, the second recombinant which were called HD-1 and No. 58pyc were target strains having the mutated hor gene and pyc gene, respectively.
[0610] (3) Lysine production test of HD-1 and No. 58pyc strains
[0611] The HD-1 strain (strain obtained by incorporating the mutation, Val59Ala, in the hor gene into the ATCC 13032 strain) and the No. 58pyc strain (strain obtained by incorporating the mutation, Pro458Ser, in the pyc gene into the lysine-producing No. 58 strain) were subjected to a culture test in a 5 1 jar fermenter by using the ATCC 13032 strain and the lysine-producing No. 58 strain respectively as a control. Thus lysine production was examined.
[0612] After culturing on BYG agar medium at 30° C. for 24 hours, each strain was inoculated into 250 ml of a seed medium (medium prepared by adding 50 g of sucrose, 40 g of corn steep liquor, 8.3 g of ammonium sulfate, 1 g of urea, 2 g of potassium dihydrogenphosphate, 0.83 g of magnesium sulfate heptahydrate, 10 mg of iron sulfate heptahydrate, 1 mg of copper sulfate pentahydrate, 10 mg of zinc sulfate heptahydrate, 10 mg of &bgr;-alanine, 5 mg of nicotinic acid, 1.5 mg of thiamin hydrochloride, and 0.5 mg of biotin to 1 liter of water, and adjusting its pH to 7.2, then to which 30 g of calcium carbonate had been added) contained in a 2 1 buffle-attached Erlenmeyer flask and cultured therein at 30° C. for 12 to 16 hours. A total amount of the seed culturing medium was inoculated into 1,400 ml of a main culture medium (medium prepared by adding 60 g of glucose, 20 g of corn steep liquor, 25 g of ammonium chloride, 2.5 g of potassium dihydrogenphosphate, 0.75 g of magnesium sulfate heptahydrate, 50 mg of iron sulfate heptahydrate, 13 mg of manganese sulfate pentahydrate, 50 mg of calcium chloride, 6.3 mg of copper sulfate pentahydrate, 1.3 mg of zinc sulfate heptahydrate, 5 mg of nickel chloride hexahydrate, 1.3 mg of cobalt chloride hexahydrate, 1.3 mg of ammonium molybdenate tetrahydrate, 14 mg of nicotinic acid, 23 mg of &bgr;-alanine, 7 mg of thiamin hydrochloride, and 0.42 mg of biotin to 1 liter of water) contained in a 5 1 jar fermenter and cultured therein at 32° C., 1 vvm and 800 rpm while controlling the pH to 7.0 with aqueous ammonia. When glucose in the medium had been consumed, a glucose feeding solution (medium prepared by adding 400 g glucose and 45 g of ammonium chloride to 1 liter of water) was continuously added. The addition of feeding solution was carried out at a controlled speed so as to maintain the dissolved oxygen concentration within a range of 0.5 to 3 ppm. After culturing for 29 hours, the culture was terminated. The cells were separated from the culture medium by centrifugation and then L-lysine hydrochloride in the supernatant was quantified by high performance liquid chromatography (HPLC). The results are shown in Table 2 below. 2 TABLE 2 Strain L-Lysine hydrochloride yield (g/l) ATCC 13032 0 HD-1 8 No. 58 45 No. 58pyc 51
[0613] As is apparent from the results shown in Table 2, the lysine productivity was improved by introducing the mutation, Val59Ala, in the hom gene or the mutation, Pro458Ser, in the pyc gene. Accordingly, it was found that the mutations are both effective mutations relating to the production of lysine. Strain, AHP-3, in which the mutation, Val59Ala, in the hom gene and the mutation, Pro458Ser, in the pyc gene have been introduced into the wild type ATCC 13032 strain together with the mutation, Thr33lIle in the lysC gene has been deposited on Dec. 5, 2000, in National Institute of Bioscience and Human Technology, Agency of Industrial Science and Technology (Higashi 1-1-3, Tsukuba-shi, Ibaraki, Japan) as FERM BP-7382.
EXAMPLE 3[0614] Reconstruction of lysine-producing strain based on genome information
[0615] The lysine-producing mutant B-6 strain (Appl. Microbiol. Biotechnol., 32:269-273 (1989)), which has been constructed by multiple round random mutagenesis with NTG and screening from the wild type ATCC 13032 strain, produces a remarkably large amount of lysine hydrochloride when cultured in a jar at 32° C. using glucose as a carbon source. However, since the fermentation period is long, .the production rate is less than 2.1 g/l/h. Breeding to reconstitute only effective mutations relating to the production of lysine among the estimated at least 300 mutations introduced into the B-6 strain in the wild type ATCC 13032 strain was performed.
[0616] (1) Identification of mutation point and effective mutation by comparing the gene nucleotide sequence of the B-6 strain with that of the ATCC 13032 strain
[0617] As described above, the nucleotide sequences of genes derived from the B-6 strain were compared with the corresponding nucleotide sequences of the ATCC 13032 strain genome represented by SEQ ID NOS:1 to 3501 and analyzed to identify many mutation points accumulated in the chromosome of the B-6 strain. Among these, a mutation, Val59lAla, in hor, a mutation, Thr311Ile, in lysC, a mutation, Pro458Ser, in pyc and a mutation, Ala2l3Thr, in zwf were specified as effective mutations relating to the production of lysine. Breeding to reconstitute the 4 mutations in the wild type strain and for constructing of an industrially important lysine-producing strain was carried out according to the method shown below.
[0618] (2) Construction of plasmid for gene replacement having mutated gene
[0619] The plasmid for gene replacement, pChom59, having the mutated hom gene and the plasmid for gene replacement, pCpyc458, having the mutated pyc gene were prepared in the above Example 2 (2). Plasmids for gene replacement having the mutated lysC and zvf were produced as described below.
[0620] The lysC and zwf having mutation points were amplified by PCR, and inserted into a plasmid for gene replacement, pCES30, according to the AT cloning method described in Example 2(2) (Bio Experiment Illustrated, Vol. 3).
[0621] Separately, chromosomal DNA was prepared from the lysine-producing B-6 strain according to the above method of Saito et al. Using the chromosomal DNA as a template, PCR was carried out with Pfu turbo DNA polymerase (manufactured by Stratagene). In the mutated lysC gene, the DNAs having the nucleotide sequences represented by SEQ ID NOS:7006 and 7007 were used as the primer set. In the mutated gene, the DNAs having the nucleotide sequences represented by SEQ ID NOS:7008 and 7009 as the primer set. The resulting PCR product was subjected to agarose gel electrophoresis, and extracted and purified using GENEGLEAN Kit (manufactured by BIO 101). Then, the PCR product was allowed to react in the presence of Taq DNA polymerase (manufactured by Roche Diagnostics) and DATP at 72° C. for 10 minutes so that a nucleotide, adenine (A), was added to the 3′-end.
[0622] The above pCES30 T vector fragment and the mutated lysC gene (1.5 kb) or mutated zwf gene (2.3 kb) to which the nucleotide A had been added of the PCR product were concentrated by extraction with phenol/chloroform and precipitation with ethanol, and then ligated using Ligation Kit ver. 2. The ligation products were introduced into the ATCC 13032 strain according to the electroporation method, and cultured on BYG agar medium containing 25 &mgr;g/ml kanamycin at 30° C. for 2 days to obtain kanamycin-resistant transformants. Each of the resulting transformants was cultured overnight in BYG liquid medium containing 25 &mgr;g/ml kanamycin, and a plasmid was extracted from the culturing solution medium according to the alkali SDS method. As a result of digestion analysis using restriction enzymes, it was confirmed that the plasmid had a structure in which the 1.5 kb or 2.3 kb DNA fragment had been inserted into pCES30. The plasmids thus constructed were named respectively pClysC311 and pCzwf2l3.
[0623] (3) Introduction of mutation, Thr3llIle, in lysC into one point mutant HD-1
[0624] Since the one mutation point mutant HD-1 in which the mutation, Val59Ala, in hom was introduced into the wild type ATCC 13032 strain had been obtained in Example 2(2), the mutation, Thr3llIle, in lysC was introduced into the HD-1 strain using pClysC311 produced in the above (2) according to the gene replacement method described in Example 2(2). PCR was carried out using chromosomal DNA of the resulting strain and, as the primer set, DNAs having the nucleotide sequences represented by SEQ ID NOS:7006 and 7007 in the same manner as in Example 2(2). As a result of the fact that the nucleotide sequence of the PCR product was determined in the usual manner, it was confirmed that the strain which was named AHD-2 was a two point mutant having the mutated lysC gene in addition to the mutated hom gene.
[0625] (4) Introduction of mutation, Pro458Ser, in pyc into two point mutant AHD-2
[0626] The mutation, Pro458Ser, in pyc was introduced into the AHD-2 strain using the pCpyc458 produced in Example 2(2) by the gene replacement method described in Example 2 (2). PCR was carried out using chromosomal DNA of the resulting strain and, as the primer set, DNAs having the nucleotide sequences represented by SEQ ID NOS:7004 and 7005 in the same manner as in Example 2 (2). As a result of the fact that the nucleotide sequence of the PCR product was determined in the usual manner, it was confirmed that the strain which was named AHD-3 was a three point mutant having the mutated pyc gene in addition to the mutated hom gene and lysC gene.
[0627] (5) Introduction of mutation, Ala2l3Thr, in zwf into three point mutant AHP-3
[0628] The mutation, Ala213Thr, in zwf was introduced into the AHP-3 strain using the pCzwf458 produced in the above (2) by the gene replacement method described in Example 2(2). PCR was carried out using chromosomal DNA of the resulting strain and, as the primer set, DNAs having the nucleotide sequences represented by SEQ ID NOS:7008 and 7009 in the same manner as in Example 2(2). As a result of the fact that the nucleotide sequence of the PCR product was determined in the usual manner, it was confirmed that the strain which was named APZ-4 was a four point mutant having the mutated zwf gene in addition to the mutated hom gene, lysC gene and pyc gene.
[0629] (6) Lysine production test on HD-1, AHD-2, AHP-3 and APZ-4 strains
[0630] The HD-1, AHD-2, AHP-3 and APZ-4 strains obtained above were subjected to a culture test in a 5 1 jar fermenter in accordance with the method of Example 2(3).
[0631] Table 3 shows the results. 3 TABLE 3 L-Lysine hydrochloride Productivity Strain (g/l) (g/l/h) HD-1 8 0.3 AHD-2 73 2.5 AHP-3 80 2.8 APZ-4 86 3.0
[0632] Since the lysine-producing mutant B-6 strain which has been bred based on the random mutation and selection shows a productivity of less than 2.1 g/l/h, the APZ-4 strain showing a high productivity of 3.0 g/l/h is useful in industry.
[0633] (7) Lysine fermentation by APZ-4 strain at high temperature
[0634] The APZ-4 strain, which had been reconstructed by introducing 4 effective mutations into the wild type strain, was subjected to the culturing test in a 5 1 jar fermenter in the same manner as in Example 2 (3), except that the culturing temperature was changed to 40° C.
[0635] The results are shown in Table 4. 4 TABLE 4 Temperature L-Lysine hydrochloride Productivity (° C.) (g/l) (g/l/h) 32 86 3.0 40 95 3.3
[0636] As is apparent from the results shown in Table 4, the lysine hydrochloride titer and productivity in culturing at a high temperature of 40° C. comparable to those at 32° C. were obtained. In the mutated and bred lysine-producing B-6 strain constructed by repeating random mutation and selection, the growth and the lysine productivity are lowered at temperatures exceeding 34° C. so that lysine fermentation cannot be carried out, whereas lysine fermentation can be carried out using the APZ-4 strain at a high temperature of 40° C. so that the load of cooling is greatly reduced and it is industrially useful. The lysine fermentation at high temperatures can be achieved by reflecting the high temperature adaptability inherently possessed by the wild type strain on the APZ-4 strain.
[0637] As demonstrated in the reconstruction of the lysine-producing strain, the present invention provides a novel breeding method effective for eliminating the problems in the conventional mutants and acquiring industrially advantageous strains. This methodology which reconstitutes the production strain by reconstituting the effective mutation is an approach which is efficiently carried out using the nucleotide sequence information of the genome disclosed in the present invention, and its effectiveness was found for the first time in the present invention.
EXAMPLE 4[0638] Production of DNA microarray and use thereof
[0639] A DNA microarray was produced based on the nucleotide sequence information of the ORF deduced from the full nucleotide sequences of Corynebacterium glutamicum ATCC 12032 using software, and genes of which expression is fluctuated depending on the carbon source during culturing were searched.
[0640] (1) Production of DNA microarray
[0641] chromosomal DNA was prepared from Corynebacterium glutamicum ATCC 13032 by the method of Saito et al. (Biochem. Biophys. Acta, 72:619 (1963)). Based on 24 genes having the nucleotide sequences represented by SEQ ID NOS:207, 3433, 281, 3435, 3439, 765, 3445, 1226, 1229, 3448, 3451, 3453, 3455, 1743, 3470, 2132, 3476, 3477, 3485, 3488, 3489, 3494, 3496, and 3497 from the ORFs shown in Table 1 deduced from the full genome nucleotide sequence of Corynebacterium glutamicum ATCC 13032 using software and the nucleotide sequence of rabbit globin gene (GenBank Accession No. V00882) used as an internal standard, oligo DNA primers for PCR amplification represented by SEQ ID NOS:7010 to 7059 targeting the nucleotide sequences of the genes were synthesized in a usual manner.
[0642] As the oligo DNA primers used for the PCR,
[0643] DNAs having the nucleotide sequence represented by SEQ ID NOS:7010 and 7011 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:207,
[0644] DNAs having the nucleotide sequence represented by SEQ ID NOS:7012 and 7013 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3433,
[0645] DNAs having the nucleotide sequence represented by SEQ ID NOS:7014 and 7015 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:281,
[0646] DNAs having the nucleotide sequence represented by SEQ ID NOS:7016 and 7017 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3435,
[0647] DNAs having the nucleotide sequence represented by SEQ ID NOS:7018 and 7019 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID No:3439,
[0648] DNAs having the nucleotide sequence represented by SEQ ID NOS:7020 and 7021 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:765,
[0649] DNAs having the nucleotide sequence represented by SEQ ID NOS:7022 and 7023 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3445,
[0650] DNAs having the nucleotide sequence represented by SEQ ID NOS:7024 and 7025 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:1226,
[0651] DNAs having the nucleotide sequence represented by SEQ ID NOS:7026 and 7027 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:1229,
[0652] DNAs having the nucleotide sequence represented by SEQ ID NOS:7028 and 7029 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3448,
[0653] DNAs having the nucleotide sequence represented by SEQ ID NOS:7030 and 7031 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3451,
[0654] DNAs having the nucleotide sequence represented by SEQ ID NOS:7032 and 7033 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3453,
[0655] DNAs having the nucleotide sequence represented by SEQ ID NOS:7034 and 7035 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3455,
[0656] DNAs having the nucleotide sequence represented by SEQ ID NOS:7036 and 7037 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:1743,
[0657] DNAs having the nucleotide sequence represented by SEQ ID NOS:7038 and 7039 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3470,
[0658] DNAs having the nucleotide sequence represented by SEQ ID NOS:7040 and 7041 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:2132,
[0659] DNAs having the nucleotide sequence represented by SEQ ID NOS:7042 and 7043 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3476,
[0660] DNAs having the nucleotide sequence represented by SEQ ID NOS:7044 and 7045 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3477,
[0661] DNAs having the nucleotide sequence represented by SEQ ID NOS:7046 and 7047 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3485,
[0662] DNAs having the nucleotide sequence represented by SEQ ID NOS:7048 and 7049 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3488,
[0663] DNAs having the nucleotide sequence represented by SEQ ID NOS:7050 and 7051 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3489,
[0664] DNAs having the nucleotide sequence represented by SEQ ID NOS:7052 and 7053 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3494,
[0665] DNAs having the nucleotide sequence represented by SEQ ID NOS:7054 and 7055 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3496,
[0666] DNAs having the nucleotide sequence represented by SEQ ID NOS:7056 and 7057 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3497, and
[0667] DNAs having the nucleotide sequence represented by SEQ ID NOS:7058 and 7059 were used for the amplification of the DNA having the nucleotide sequence of the rabbit globin gene,
[0668] as the respective primer set.
[0669] The PCR was carried for 30 cycles with each cycle consisting of 15 seconds at 95° C. and 3 minutes at 68° C. using a thermal cycler (GeneAmp PCR system 9600, manufactured by Perkin Elmer), TaKaRa EX-Taq (manufactured by Takara Shuzo), 100 ng of the chromosomal DNA and the buffer attached to the TaKaRa Ex-Taq reagent. In the case of the rabbit globin gene, a single-stranded cDNA which had been synthesized from rabbit globin mRNA (manufactured by Life Technologies) according to the manufacture's instructions using a reverse transcriptase RAV-2 (manufactured by Takara Shuzo). The PCR product of each gene thus amplified was subjected to agarose gel electrophoresis and extracted and purified using QIAquick Gel Extraction Kit (manufactured by QIAGEN). The purified PCR product was concentrated by precipitating it with ethanol and adjusted to a concentration of 200 ng/&mgr;l. Each PCR product was spotted on a slide glass plate (manufactured by Matsunami Glass) having MAS coating in 2 runs using GTMASS SYSTEM (manufactured by Nippon Laser & Electronics Lab.) according to the manufacture's instructions.
[0670] (2) Synthesis of fluorescence labeled cDNA
[0671] The ATCC 13032 strain was spread on BY agar medium (medium prepared by adding 20 g of peptone (manufactured by Kyokuto Pharmaceutical), 5 g of yeast extract (manufactured by Difco), and 16 g of Bactoagar (manufactured by Difco) to in 1 liter of water and adjusting its pH to 7.2) and cultured at 30° C. for 2 days. Then, the cultured strain was further inoculated into 5 ml of BY liquid medium and cultured at 30° C. overnight. Then, the cultured strain was further inoculated into 30 ml of a minimum medium (medium prepared by adding 5 g of ammonium sulfate, 5 g of urea, 0.5 g of monopotassium dihydrogenphosphate, 0.5 g of dipotassium monohydrogenphosphate, 20.9 g of morpholinopropanesulfonic acid, 0.25 g of magnesium sulfate heptahydrate, 10 mg of calcium chloride dihydrate, 10 mg of manganese sulfate monohydrate, 10 mg of ferrous sulfate heptahydrate, 1 mg of zinc sulfate heptahydrate, 0.2 mg copper sulfate, and 0.2 mg biotin to 1 liter of water, and adjusting its pH to 6.5) containing 110 mmol/l glucose or 200 mmol/l ammonium acetate, and cultured in an Erlenmyer flask at 30° to give 1.0 of absorbance at 660 nm. After the cells were prepared by centrifuging at 4° C. and 5,000 rpm for 10 minutes, total RNA was prepared from the resulting cells according to the method of Bormann et al. (Molecular Microbiology, 6:317-326 (1992)). To avoid contamination with DNA, the RNA was treated with DnaseI (manufactured by Takara Shuzo) at 37° C. for 30 minutes and then further purified using Qiagen RNeasy MiniKit (manufactured by QIAGEN) according to the manufacture's instructions. To 30 fg of the resulting total RNA, 0.6 &mgr;l of rabbit globin MRNA (50 ng/&mgr;l, manufactured by Life Technologies) and 1 &mgr;l of a random 6 mer primer (500 ng/&mgr;l, manufactured by Takara Shuzo) were added for denaturing at 65° C. for 10 minutes, followed by quenching on ice. To the resulting solution, 6 &mgr;l of a buffer attached to SuperScript II (manufactured by Lifetechnologies), 3 &mgr;l of 0.1 mol/l DTT, 1.5 &mgr;l of dNTPs (25 mmol/l DATP, 25 rmnol/l dCTP, 25 mmol/l dGTP, 10 mmol/l dTTP), 1.5 &mgr;l of Cy5-dUTP or Cy3-dUTP (manufactured by NEN) and 2 &mgr;l of SuperScript II were added, and allowed to stand at 25° C. for 10 minutes and then at 42° C. for 110 minutes. The RNA extracted from the cells using glucose as the carbon source and the RNA extracted from the cells using ammonium acetate were labeled with Cy5-dUTP and Cy3-dUTP, respectively. After the fluorescence labeling reaction, the RNA was digested by adding 1.5 &mgr;l of 1 mol/l sodium hydroxide-20 mmol/l EDTA solution and 3.0 &mgr;l of 10% SDS solution, and allowed to stand at 65° C. for 10 minutes. The two cDNA solutions after the labeling were mixed and purified using Qiagen PCR purification Kit (manufactured by QIAGEN) according to the manufacture's instructions to give a volume of 10 &mgr;l.
[0672] (3) Hybridization
[0673] UltraHyb (110 &mgr;l) (manufactured by Ambion) and the fluorescence-labeled CDNA solution (10 &mgr;l) were mixed and subjected to hybridization and the subsequent washing of slide glass using GeneTAC Hybridization Station (manufactured by Genomic Solutions) according to the manufacture's instructions. The hybridization was carried out at 50° C., and the washing was carried out at 25° C.
[0674] (4) Fluorescence analysis
[0675] The fluorescence amount of each DNA array having the fluorescent CDNA hybridized therewith was measured using ScanArray 4000 (manufactured by GSI Lumonics).
[0676] Table 5 shows the Cy3 and Cy5 signal intensities of the genes having been corrected on the basis of the data of the rabbit globin used as the internal standard and the Cy3/Cy5 ratios. 5 TABLE 5 SEQ ID NO Cy3 intensity Cy5 intensity Cy3/Cy5 207 5248 3240 1.62 3433 2239 2694 0.83 281 2370 2595 0.91 3435 2566 2515 1.02 3439 5597 6944 0.81 765 6134 4943 1.24 3455 1169 1284 0.91 1226 1301 1493 0.87 1229 1168 1131 1.03 3448 1187 1594 0.74 3451 2845 3859 0.74 3453 3498 1705 2.05 3455 1491 1144 1.30 1743 1972 1841 1.07 3470 4752 3764 1.26 2132 1173 1085 1.08 3476 1847 1420 1.30 3477 1284 1164 1.10 3485 4539 8014 0.57 3488 34289 1398 24.52 3489 43645 1497 29.16 3494 3199 2503 1.28 3496 3428 2364 1.45 3497 3848 3358 1.15
[0677] The ORF function data estimated by using software were searched for SEQ ID NOS:3488 and 3489 showing remarkably strong Cy3 signals. As a result, it was found that SEQ ID NOS:3488 and 3489 are a maleate synthase gene and an isocitrate lyase gene, respectively. It is known that these genes are transcriptionally induced by acetic acid in Corynebacterium glutamicum (Archives of Microbiology, 168:262-269 (1997)).
[0678] As described above, a gene of which expression is fluctuates could be discovered by synthesizing appropriate oligo DNA primers based on the ORF nucleotide sequence information deduced from the full genomic nucleotide sequence information of Corynebacterium glutamicum ATCC 13032 using software, amplifying the nucleotide sequences of the gene using the genome DNA of Corynebacterium glutamicum as a template in the PCR reaction, and thus producing and using a DNA microarray.
[0679] This Example shows that the expression amount can be analyzed using a DNA microarray in the 24 genes. On the other hand, the present DNA microarray techniques make it possible to prepare DNA microarrays having thereon several thousand gene probes at once. Accordingly, it is also possible to prepare DNA microarrays having thereon all of the ORF gene probes deduced from the full genomic nucleotide sequence of Corynebacterium glutamicum ATCC 13032 determined by the present invention, and analyze the expression profile at the total gene level of Corynebacterium glutamicum using these arrays.
EXAMPLE 5[0680] Homology search using Corynebacterium glutamicum genome sequence
[0681] (1) Search of adenosine deaminase
[0682] The amino acid sequence (ADDECOLI) of Escherichia coli adenosine deaminase was obtained from Swiss-prot Database as the amino acid sequence of the protein of which function had been confirmed as adenosine deaminase (EC3.5.4.4). By using the full length of this amino acid sequence as a query, a homology search was carried out on a nucleotide sequence database of the genome sequence of Corynebacterium glutamicum or a database of the amino acids in the ORF region deduced from the genome sequence using FASTA program (Proc. Natl. Acad. Sci. ISA, 85:2444-2448 (1988)). A case where E-value was le−10 or less was judged as being significantly homologous. As a result, no sequence significantly homologous with the Escherichia coli adenosine deaminase was found in the nucleotide sequence database of the genome sequence of Corynebacterium glutamicum or the database of the amino acid sequences in the ORF region deduced from the genome sequence. Based on these results, it is assumed that Corynebacterium glutamicum contains no ORF having adenosine deaminase activity and thus has no activity of converting adenosine into inosine.
[0683] (2) Search of glycine cleavage enzyme
[0684] The sequences (GCSP_ECOLI, GCST_ECOLI and GCSH_ECOLI) of glycine decarboxylase, aminomethyl transferase and an aminomethyl group carrier each of which is a component of Escherichia coli glycine cleavage enzyme as the amino acid sequence of the protein, of which function had been confirmed as glycine cleavage enzyme (EC2.1.2.10), were obtained from Swiss-prot Database.
[0685] By using these full-length amino acid sequences as a query, a homology search was carried out on a nucleotide sequence database of the genome sequence of Corynebacterium glutamicum or a database of the ORF amino acid sequences deduced from the genome sequence using FASTA program. A case where E-value was le−10 or less was judged as being significantly homologous. As a result, no sequence significantly homologous with the glycine decarboxylase, the aminomethyl transferase or the aminomethyl group carrier each of which is a component of Escherichia coil glycine cleavage enzyme, was found in the nucleotide sequence database of the genome sequence of Corynebacterium glutamicum or the database of the ORF amino acid sequences estimated from the genome sequence. Based on these results, it is assumed that Corynebacterium glutamicum contains no ORF having the activity of glycine decarboxylase, aminomethyl transferase or the aminomethyl group carrier and thus has no activity of the glycine cleavage enzyme.
[0686] (3) Search of IMP dehydrogenase
[0687] The amino acid sequence (IMDH ECOLI) of Escherichia coli IMP dehydrogenase as the amino acid sequence of the protein, of which function had been confirmed as IMP dehydrogenase (EC1.1.1.205), was obtained from Swiss-prot Database. By using the full length of this amino acid sequence as a query, a homology search was carried out on a nucleotide sequence database of the genome sequence of Corynebacterium glutamicum or a database of the ORF amino acid sequences predicted from the genome sequence using FASTA program. A case where E-value was le−10 or less was judged as being significantly homologous. As a result, the amino acid sequences encoded by two ORFS, namely, an ORF positioned in the region of the nucleotide sequence No. 615336 to 616853 (or ORF having the nucleotide sequence represented by SEQ ID NO:672) and another ORF positioned in the region of the nucleotide sequence No. 616973 to 618094 (or ORF having the nucleotide sequence represented by SEQ ID NO:674) were significantly homologous with the ORFs of Escherichia coli IMP dehydrogenase. By using the above-described predicted amino acid sequence as a query in order to examine the similarity of the amino acid sequences encoded by the ORFs with IMP dehydrogenases of other organisms in greater detail, a search was carried out on GenBank (http://www.ncbi.nlm.nih.gov/) nr-aa database (amino acid sequence database constructed on the basis of GenBankCDS translation products, PDB database, Swiss-Prot database, PIR database, PRF database by eliminating duplicated registrations) using BLAST program. As a result, both of the two amino acid sequences showed significant homologies with IMP dehdyrogenases of other organisms and clearly higher homologies with IMP dehdyrogenases than with amino acid sequences of other proteins, and thus, it was assumed that the two ORFs would function as IMP dehydrogenase. Based on these results, it was therefore assumed that Corynebacterium glutamicum has two ORFs having the IMP dehydrogenase activity.
EXAMPLE 6[0688] Proteome analysis of proteins derived from Corynebacterium glutamicum
[0689] (1) Preparations of proteins derived from Corynebacterium glutamicum ATCC 13032, FERM BP-7134 and FERM BP-158
[0690] Culturing tests of Corynebacterium glutamicum ATCC 13032 (wild type strain), Corynebacterium glutamicum FERM BP-7134 (lysine-producing strain) and Corynebacterium glutamicum (FERM BP-158, lysine-highly producing strain) were carried out in a 5 1 jar fermenter according to the method in Example 2(3). The results are shown in Table 6. 6 TABLE 6 Strain L-Lysine yield (g/l) ATCC 13032 0 FERM BP-7134 45 FERM BP-158 60
[0691] After culturing, cells of each strain were recovered by centrifugation. These cells were washed with Tris-HCl buffer (10 mmol/l Tris-HCl, pH 6.5, 1.6 mg/ml protease inhibitor (COMPLETE; manufactured by Boehringer Mannheim)) three times to give washed cells which could be stored under freezing at −80° C. The freeze-stored cells were thawed before use, and used as washed cells.
[0692] The washed cells described above were suspended in a disruption buffer (10 mmol/l Tris-HCl, pH 7.4, 5 mmol/l magnesium chloride, 50 mg/l RNase, 1.6 mg/ml protease inhibitor (COMPLETE:manufactured by Boehringer Mannheim)), and disrupted with a disruptor (manufactured by Brown) under cooling. To the resulting disruption solution, DNase was added to give a concentration of 50 mg/l, and allowed to stand on ice for 10 minutes. The solution was centrifuged (5,000×g, 15 minutes, 4° C.) to remove the undisrupted cells as the precipitate, and the supernatant was recovered.
[0693] To the supernatant, urea was added to give a concentration of 9 mol/l, and an equivalent amount of a lysis buffer (9.5 mol/l urea, 2% NP-40, 2% Ampholine, 5% mercaptoethanol, 1.6 mg/ml protease inhibitor (COMPLETE; manufactured by Boehringer Mannheim) was added thereto, followed by thoroughly stirring at room temperature for dissolving.
[0694] After being dissolved, the solution was centrifuged at 12,000×g for 15 minutes, and the supernatant was recovered.
[0695] To the supernatant, ammonium sulfate was added to the extent of 80% saturation, followed by thoroughly stirring for dissolving.
[0696] After being dissolved, the solution was centrifuged (16,000×g, 20 minutes, 4° C.), and the precipitate was recovered. This precipitate was dissolved in the lysis buffer again and used in the subsequent procedures as a protein sample. The protein concentration of this sample was determined by the method for quantifying protein of Bradford.
[0697] (2) Separation of protein by two dimensional electrophoresis
[0698] The first dimensional electrophoresis was carried out as described below by the isoelectric electrophoresis method.
[0699] A molded dry IPG strip gel (pH 4-7, 13 cm, Immobiline DryStrips; manufactured by Amersham Pharmacia Biotech) was set in an electrophoretic apparatus (Multiphor II or IPGphor; manufactured by Amersham Pharmacia Biotech) and a swelling solution (9 mol/l urea, 0.5% Triton X-100, 0.6% dithiothreitol, 0.5%W mpholine, pH 3-10) was packed therein, and the gel was allowed to stand for swelling 12 to 16 hours.
[0700] The protein sample prepared above was dissolved in a sample solution (9 mol/l urea, 2% CHAPS, 1% dithiothreitol, 2% Ampholine, pH 3-10), and then about 100 to 500 &mgr;g (in terms of protein) portions thereof were taken and added to the swollen IPG strip gel.
[0701] The electrophoresis was carried out in the 4 steps as defined below under controlling the temperature to 20° C.:
[0702] step 1:1 hour under a gradient mode of 0 to 500v;
[0703] step 2:1 hour under a gradient mode of 500 to 1,000 V;
[0704] step 3:4 hours under a gradient mode of 1,000 to 8,000 V; and
[0705] step 4:1 hour at a constant voltage of 8,000 V.
[0706] After the isoelectric electrophoresis, the IPG strip gel was put off from the holder and soaked in an equilibration buffer A (50 mmol/l Tris-HCl, pH 6.8, 30% glycerol, 1% SDS, 0.25t dithiothreitol) for 15 minutes and another equilibration buffer B (50 nmol/l Tris-HCl, pH 6.8, 6 mol/l urea, 30t glycerol, 1% SDS, 0.45% iodo acetamide) for 15 minutes to sufficiently equilibrate the gel.
[0707] After the equilibrium, the IPG strip gel was lightly rinsed in an SDS electrophoresis buffer (1.4% glycine, 0.1% SDS, 0.3% Tris-HCl, pH 8.5), and the second dimensional electrophoresis depending on molecular weight was carried out as described below to separate the proteins.
[0708] Specifically, the above IPG strip gel was closely placed on 14% polyacrylamide slub gel (14% polyacrylamide, 0.37% bisacrylamide, 37.5 mmol/l Tris-HCl, pH 8.8, 0.1% SDS, 0.1% TEMED, 0.1% ammonium persulfate) and subjected to electrophoresis under a constant voltage of 30 mA at 20° C for 3 hours to separate the proteins.
[0709] (3) Detection of protein spot
[0710] Coomassie staining was performed by the method of Gorg et al. (Electrophoresis, 9:531-546 (1988)) for the slub gel after the second dimensional electrophoresis. Specifically, the slub gel was stained under shaking at 25° C. for about 3 hours, the excessive coloration was removed with a decoloring solution, and the gel was thoroughly washed with distilled water.
[0711] The results are shown in FIG. 2. The proteins derived from the ATCC 13032 strain (FIG. 2A), FERM BP-7134 strain (FIG. 2B) and FERM BP-158 strain (FIG. 2C) could be separated and detected as spots.
[0712] (4) In-gel digestion of detected protein spot
[0713] The detected spots were each cut out from the gel and transferred into siliconized tube, and 400 &mgr;l of 100 mmol/l ammonium bicarbonate:acetonitrile solution (1:1, v/v) was added thereto, followed by shaking overnight and freeze-dried as such. To the dried gel, 10 &mgr;l of a lysylendopeptidase (LysC) solution (manufactured by WAKO, prepared with 0.1% SDS-containing 50 mmol/l ammonium bicarbonate to give a concentration of 100 ng/&mgr;l) was added and the gel was allowed to stand for swelling at 0° C. for 45 minutes, and then allowed to stand at 37° C. for 16 hours. A.After removing the LysC solution, 20 &mgr;l of an extracting solution (a mixture of 60% acetonitrile and 5% formic acid) was added, followed by ultrasonication at room temperature for 5 minutes to disrupt the gel. After the disruption, the extract was recovered by centrifugation (12,000 rpm, 5 minutes, room temperature). This operation was repeated twice to recover the whole extract. The recovered extract was concentrated by centrifugation in vacuo to halve the liquid volume. To the concentrate, 20 &mgr;l of 0.1% trifluoroacetic acid was added, followed by thoroughly stirring, and the mixture was subjected to desalting using ZipTip (manufactured by Millipore). The protein absorbed on the carriers of ZipTip was eluted with 5 &mgr;l of &agr;-cyano-4-hydroxycinnamic acid for use as a sample solution for analysis.
[0714] (5) Mass spectrometry and amino acid sequence analysis of protein spot with matrix assisted laser desorption ionization time of flight mass spectrometer (MALDI-TOFMS)
[0715] The sample solution for analysis was mixed in the equivalent amount with a solution of a peptide mixture for mass calibration (300 nmol/l Angiotensin II, 300 nmol/l Neurotensin, 150 nmol/l ACTHclip 18-39, 2.3 &mgr;mol/l bovine insulin B chain), and 1 &mgr;l of the obtained solution was spotted on a stainless probe and crystallized by spontaneously drying.
[0716] As measurement instruments, REFLEX MALDI-TOF mass spectrometer (manufactured by Bruker) and an N2 laser (337 nm) were used in combination.
[0717] The analysis by PMF (peptide-mass finger printing) was carried out using integration spectra data obtained by measuring 30 times at an accelerated voltage of 19.0 kV and a detector voltage of 1.50 kV under reflector mode conditions. Mass calibration was carried out by the internal standard method.
[0718] The PSD (post-source decay) analysis was carried out using integration spectra obtained by successively altering the reflection voltage and the detector voltage at an accelerated voltage of 27.5 kV.
[0719] The masses and amino acid sequences of the peptide fragments derived from the protein spot after digestion were thus determined.
[0720] (6) Identification of protein spot
[0721] From the amino acid sequence information of the digested peptide fragments derived from the protein spot obtained in the above (5), ORFs corresponding to the protein were searched on the genome sequence database of Corynebacterium glutamicum ATCC 13032 as constructed in Example 1 to identify the protein.
[0722] The identification of the protein was carried out using MS-Fit program and MS-Tag program of intranet protein prospector.
[0723] (a) Search and identification of gene encoding high-expression protein
[0724] In the proteins derived from Corynebacterium glutamicum ATCC 13032 showing high expression amounts in CBB-staining shown in FIG. 2A, the proteins corresponding to Spots-1, 2, 3, 4 and 5 were identified by the above method.
[0725] As a result, it was found that Spot-1 corresponded to enolase which was a protein having the amino acid sequence of SEQ ID NO:4585; Spot-2 corresponded to phosphoglycelate kinase which was a protein having the amino acid sequence of SEQ ID NO:5254; Spot-3 corresponded to glyceraldehyde-3-phosphate dehydrogenase which was a protein having the amino acid sequence represented by SEQ ID NO:5255; Spot-4 corresponded to fructose bis-phosphatg aldolase which was a protein having the amino acid sequence represented by SEQ ID NO:6543; and Spot-5 corresponded to triose phosphate isomerase which was a protein having the amino acid sequence represented by SEQ ID NO:5252.
[0726] These genes, represented by SEQ ID NOS:1085, 1754, 1775, 3043 and 1752 encoding the proteins corresponding to Spots-l, 2, 3, 4 and 5, respectively, encoding the known proteins are important in the central metabolic pathway for maintaining the life of the microorganism. Particularly, it is suggested that the genes of Spots-2, 3 and 5 form an operon and a high-expretsion promoter is encoded in the upstream thereof (J. of Bacteriol., 174:6067-6086 (1992)).
[0727] Also, the protein corresponding to Spot-9 in FIG. 2 was identified in the same manner as described above, and it was found that Spot-9 was an elongation factor Tu which was a protein having the amino acid sequence represented by SEQ ID No:6937, and that the protein was encoded by DNA having the nucleotide sequence represented by SEQ ID NO:3437.
[0728] Based on these results, the proteins having high expression level were identified by proteome analysis using the genome sequence database of Corynebacterium glutamicum constructed in Example 1. Thus, the nucleotide sequences of the genes encoding the proteins and the nucleotide sequences upstream thereof could be searched simultaneously. Accordingly, it is shown that nucleotide sequences having a function as a high-expression promoter can be efficiently selected.
[0729] (b) Search and identification of modified protein
[0730] Among the proteins derived from Corynebacterium glutamicum FERM BP-7134 shown in FIG. 2B, Spots-6, 7 and 8 were identified by the above method. As a result, these three spots all corresponded to catalase which was a protein having the amino acid sequence represented by SEQ ID NO:3785.
[0731] Accordingly, all of Spots-6, 7 and 8 detected as spots differing in isoelectric mobility were all products derived from a catalase gene having the nucleotide sequence represented by SEQ ID NQ:285, Accordingly, it is shown that the catalase derived from Corynebacterium glutamicum FERM BP-7134 was modified after the translation.
[0732] Based on these results, it is confirmed that various modified proteins can be efficiently searched by proteome analysis using the genome sequence database of Coaebacteriium glutamicum constructed in Example 1.
[0733] (c) Search and identification of expressed protein effective in lysine production
[0734] It was found out that in FIG. 2A (ATCC 13032:wild type strain), FIG. 2B ( FRM BP-7134:lysine-producing strain) and FIG. 2C (FF.M BP-158:lysine-highly producing strain), the catalase corresponding to Spot-8 and the elongation factor Tu corresponding to Spot-9 as identified above showed the higher expression level with an increase in the lysine productivity.
[0735] Based on these results, it was found that hopeful mutated proteins can be efficiently searched and identified in breeding aiming at strengthening the productivity of a target product by the proteome analysis using the genome sequence database of Corynebacterium glutamicum constructed in Example 1.
[0736] Moreover, useful mutation points of useful mutants can be easily specified by searching the nucleotide sequences (nucleotide sequences of promoter, ORF, or the like) relating to the identified proteins using the above database and using primers designed on the basis of the sequences. As a result of the fact that the mutation points are specified, industrially useful mutants which have the useful mutations or other useful mutations derived therefrom can be easily bred.
[0737] While the invention has been described in detail and with reference to specific embodiments thereof, it will be apparent to one of skill in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof. All references cited herein are incorporated in their entirety
Claims
1. A method for at least one of the following:
- (A) identifying a mutation point of a gene derived from a mutant of a coryneform bacterium,
- (B) measuring an expression amount of a gene derived from a coryneform bacterium,
- (C) analyzing an expression profile of a gene derived from a coryneform bacterium,
- (D) analyzing expression patterns of genes derived from a coryneform bacterium, or
- (E) identifying a gene homologous to a gene derived from a coryneform bacterium,
- said method comprising:
- (a) producing a polynucleotide array by adhering to a solid support at least two polynucleotides selected from the group consisting of first polynucleotides comprising the nucleotide sequence represented by any one of SEQ ID NOS:1 to 3501, second polynucleotides which hybridize with the first polynucleotides under stringent conditions, and third polynucleotides comprising a sequence of 10 to 200 continuous bases of the first or second polynucleotides,
- (b) incubating the polynucleotide array with at least one of a labeled polynucleotide derived from a coryneform bacterium, a labeled polynucleotide derived from a mutant of the coryneform bacterium or a labeled polynucleotide to be examined, under hybridization conditions,
- (c) detecting any hybridization, and
- (d) analyzing the result of the hybridization.
2. The method according to claim 1, wherein the coryneform bacterium is a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterinum.
3. The method according to claim 2, wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacteriunm acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacteriunm melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
4. The method according to claim 1, wherein the polynucleotide derived from a coryneform bacterium, the polynucleotide derived from a mutant of the coryneform bacterium or the polynucleotide to be examined is a gene relating to the biosynthesis of at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof.
5. The method according to claim 1, wherein the polynucleotide to be examined is derived from Escherichia coli.
6. A polynucleotide array, comprising:
- at least two polynucleotides selected from the group consisting of first polynucleotides comprising the nucleotide sequence represented by any one of SEO ID NOS:1 to 3501, second polynucleotides which hybridize with the first polynucleotides under stringent conditions, and third polynucleotides comprising 10 to 200 continuous bases of the first or second polynucleotides, and
- a solid support adhered thereto.
7. A polynucleotide comprising the nucleotide sequence represented by SEQ ID NO:1 or a polynucleotide having a homology of at least 80% with the polynucleotide.
8. A polynucleotide comprising any one of the nucleotide sequences represented by SEQ ID NOS:2 to 3431, or a polynucleotide which hybridizes with the polynucleotide under stringent conditions.
9. A polynucleotide encoding a polypeptide having any one of the amino acid sequences represented by SEQ ID NOS:3502 to 6931, or a polynucleotide which hybridizes therewith under stringent conditions.
10. A polynucleotide which is present in the 5′ upstream or 3′ downstream of a polynucleotide comprising the nucleotide sequence of any one of SEQ ID NOS:2 to 3431 in a whole polynucleotide comprising the nucleotide sence represented by SEQ ID NO:1, and has an activity of regulating an expression of the polynucleotide.
11. A polynucleotide comprising 10 to 200 continuous bases in the nucleotide sequence of the polynucleotide of any one of claims 7 to 10, or a polynucleotide comprising a nucleotide sequence complementary to the polynucleotide comprising 10 to 200 continuous based.
12. A recombinant DNA comprising the polynucleotide of any one of claims 8 to 11.
13. A transformant comprising the polynucleotide of any one of claims 8 to 11 or the recombinant DNA of claim 12.
14. A method for producing a polypeptide, comprising:
- culturing the transformant of claim 13 in a medium to produce and accumulate a polypeptide encoded by the polynucleotide of claim 8 or 9 in the medium, and
- recovering the polypeptide from the medium.
15. A method for producing at least one of an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof, comprising:
- culturing the transformant of claim 13 in a medium to produce and accumulate at least one of an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof in the medium, and
- recovering the at least one of the amino acid, the nucleic acid, the vitamin, the saccharide, the organic acid, and analogues thereof from the medium.
16. A polypeptide encoded by a polynucleotide comprising the nucleotide sequence selected from SEQ ID NOS:2 to 3431.
17. A polypeptide comprising the amino acid sequence selected from SEQ ID NOS:3502 to 6931.
18. The polypeptide according to claim 16 or 17, wherein at least one amino acid is deleted, replaced, inserted or added, said polypeptides having an activity which is substantially the same as that of the polypeptide without said at least one amino acid deletion, replacement, insertion or addition.
19. A polypeptide comprising an amino acid sequence having a homology of at least 60% with the amino acid sequence of the polypeptide of claim 16 or 17, and having an activity which is substantially the same as that of the polypeptide.
20. An antibody which recognizes the polypeptide of any one of claims 16 to 19.
21. A polypeptide array, comprising:
- at least one polypeptide or partial fragment polypeptide selected from the polypeptides of claims 16 to 19 and partial fragment polypeptides of the polypeptides, and
- a solid support adhered thereto.
22. A polypeptide array, comprising:
- at least one antibody which recognizes a polypeptide or partial fragment polypeptide selected from the polypeptides of claims 16 to 19 and partial fragment polypeptides of the polypeptides, and
- a solid support adhered thereto.
23. A system based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
- (i) a user input device that inputs at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501, and target sequence or target structure motif information;
- (ii) a data storage device for at least temporarily storing the input information;
- (iii) a comparator that compares the at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501 with the target sequence or target structure motif information, recorded by the data storage device for screening and analyzing nucleotide sequence information which is coincident with or analogous to the target sequence or target structure motif information; and
- (iv) an output device that shows a screening or analyzing result obtained by the comparator.
24. A method based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
- (i) inputting at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501, target sequence information or target structure motif information into a user input device;
- (ii) at least temporarily storing said information;
- (iii) comparing the at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501 with the target sequence or target structure motif information; and
- (iv) screening and analyzing nucleotide sequence information which is coincident with or analogous to the target sequence or target structure motif information.
25. A system based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
- (i) a user input device that inputs at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, and target sequence or target structure motif information;
- (ii) a data storage device for at least temporarily storing the input information;
- (iii) a comparator that compares the at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target sequence or target structure motif information, recorded by the data storage device for screening and analyzing amino acid sequence information which is coincident with or analogous to the target sequence or target structure motif information; and
- (iv) an output device that shows a screening or analyzing result obtained by the comparator.
26. A method based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
- (i) inputting at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, and target sequence information or target structure motif information into a user input device;
- (ii) at least temporarily storing said information;
- (iii) comparing the at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target sequence or target structure motif information; and
- (iv) screening and analyzing amino acid sequence information which is coincident with or analogous to the target sequence or target structure motif information.
27. A system based on a computer for determining a function of a polypeptide encoded by a polynucleotide having a target nucleotide sequence derived from a coryneform bacterium, comprising the following:
- (i) a user input device that inputs at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501, function information of a polypeptide encoded by the nucleotide sequence, and target nucleotide sequence information;
- (ii) a data storage device for at least temporarily storing the input information;
- (iii) a comparator that compares the at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501 with the target nucleotide sequence information for determining a function of a polypeptide encoded by a polynucleotide having the target nucleotide sequence which is coincident with or analogous to the polynucleotide having at least one nucleotide sequence selected from SEQ ID NOS:2 to 3501; and
- (iv) an output devices that shows a function obtained by the comparator.
28. A method based on a computer for determining a function of a polypeptide encoded by a polypeptide encoded by a polynucleotide having a target nucleotide sequence derived from a coryneform bacterium, comprising the following:
- (i) inputting at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501, function information of a polypeptide encoded by the nucleotide sequence, and target nucleotide sequence information;
- (ii) at least temporarily storing said information;
- (iii) comparing the at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501 with the target nucleotide sequence information; and
- (iv) determining a function of a polypeptide encoded by a polynucleotide having the target nucleotide sequence which is coincident with or analogous to the polynucleotide having at least one nucleotide sequence selected from SEQ ID NOS;2 to 3501.
29. A system based on a computer for determining a function of a polypeptide having a target amino acid sequence derived from a coryneform bacterium, comprising the following:
- (i) a user input device that inputs at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, function information based on the amino acid sequence, and target amino acid sequence information;
- (ii) a data storing device for at least temporarily storing the input information;
- (iii) a comparator that compares the at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target amino acid sequence information for determining a function of a polypeptide having the target amino acid sequence which is coincident with or analogous to the polypeptide having at least one amino acid sequence selected from SEQ ID NOS.3502 to 7001; and
- (iv) an output device that shows a function obtained by the comparator.
30. A method based on a computer for determining a function of a polypeptide having a target amino acid sequence derived from a coryneform bacterium, comprising the following:
- (i) inputting at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, function information based on the amino acid sequence, and target amino acid sequence information;
- (ii) at least temporarily storing said information;
- (iii) comparing the at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target amino acid sequence information; and
- (iv) determining a function of a polypeptide having the target amino acid sequence which is coincident with or analogous to the polypeptide having at least one amino acid sequence selected from SEQ ID NOS:3502 to 7001.
31. The system according to any one of claims 23, 25, 27 and 29, wherein a coryneform bacterium is a microorganism of the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterinum.
32. The method according to any one of claims 24, 26, 28 and 30, wherein a coryneform bacterium is a microorganism of the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterinum.
33. The system according to claim 31, wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
34. The method according to claim 32, wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
35. A recording medium or storage device which is readable by a computer in which at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501 or function information based on the nucleotide sequence is recorded, and is usable in the system of claim 23 or 27 or the method of claim 24 or 28.
36. A recording medium or storage device which is readable by a computer in which at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 or function information based on the amino acid sequence is recorded, and is usable in the system of claim 25 or 29 or the method of claim 26 or 30.
37. The recording medium or storage device according to claim 35 or 36, which is a computer readable recording medium selected from the group consisting of a floppy disc, a hard disc, a magnetic tape, a random access memory (RAM), a read only memory (ROM), a magneto-optic disc (MO), CD-ROM, CD-R, CD-RW, DVD-ROM, DVD-RAM and DVD-RW.
38. A polypeptide having a homoserine dehydrogenase activity, comprising an amino acid sequence in which the Val residue at the 59th in the amino acid sequence of homoserine dehydrogenase derived from a coryneform bacterium is replaced with an amino acid residue other than a Val residue.
39. A polypeptide comprising an amino acid sequence in which the Val residue at the 59th position in the amino acid sequence as represented by SEQ ID NO:6952 is replaced with an amino acid residue other than a Val residue.
40. The polypeptide according to claim 38 or 39, wherein the Val residue at the 59th position is replaced with an Ala residue.
41. A polypeptide having private carboxylase activity, comprising an amino acid sequence in which the Pro residue at the 458th position in the amino acid sequence of private carboxylase derived from a coryneform bacterium is replaced with an amino acid residue other than a Pro residue.
42. A polypeptide comprising an amino acid sequence in which the Pro residue at the 458th position in the amino acid sequence represented by SEQ ID NO:4265 is replaced with an amino acid residue other than a Pro residue.
43. The polypeptide according to claim 41 or 42, wherein the Pro residue at the 458th position is replaced with a Ser residue.
44. The polypeptide according to any one of claims 38 to 43, which is derived from Corynebacterium glutamicum.
45. A DNA encoding the polypeptide of any one of claims 38 to 44.
46. A recombinant DNA comprising the DNA of claim 45.
47. A transformant comprising the recombinant DNA of claim 46.
48. A transformant comprising in its chromosome the DNA of claim 45.
49. The transformant according to claim 47 or 48, which is derived from a coryneform bacterium.
50. The transformant according to claim 49, which is derived from Corynebacterium glutamicum.
51. A method for producing L-lysine, comprising:
- culturing the transformant of any one of claims 47 to 50 in a medium to produce and accumulate L-lysine in the medium, and
- recovering the L-lysine from the culture.
52. A method for breeding a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:1 to 3431, comprising the following:
- (i) comparing a nucleotide sequence of a genome or gene of a production strain derived a coryneform bacterium which has been subjected to mutation breeding so as to produce at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogous thereof by a fermentation method, with a corresponding nucleotide sequence in SEQ ID NOS:1 to 3431;
- (ii) identifying a mutation point present in the production strain based on a result obtained by (i);
- (iii) introducing the mutation point into a coryneform bacterium which is free of the mutation point, or deleting the mutation point from a coryneform bacterium having the mutation point; and
- (iv) examining productivity by the fermentation method of the compound selected in (i) of the coryneform bacterium obtained in (iii).
53. The method according to claim 52, wherein the gene is a gene encoding an enzyme in a biosynthesis pathway or a signal transmission pathway.
54. The method according to claim 52, wherein the mutation point is a mutation point relating to a useful mutation which improves or stabilizes the productivity.
55. A method for breading a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:1 to 3431, comprising:
- (i) comparing a nucleotide sequence of a genome or gene of a production strain derived a coryneform bacterium which has been subjected to mutation breeding so as to produce at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogous thereof by a fermentation method, with a corresponding nucleotide sequence in SEQ ID NOS:1 to 3431;
- (ii) identifying a mutation point present in the production strain based on a result obtain by (i);
- (iii) deleting a mutation point from a coryneform bacterium having the mutation point; and
- (iv) examining productivity by the fermentation method of the compound selected in (i) of the coryneform bacterium obtained in (iii).
56. The method according to claim 55, wherein the gene is a gene encoding an enzyme in a biosynthesis pathway or a signal transmission pathway.
57. The method according to claim 55, wherein the mutation point is a mutation point which decreases or destabilizes the productivity.
58. A method for breeding a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:2 to 3431, comprising the following:
- (i) identifying an isozyme relating to biosynthesis of at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogous thereof, based on the nucleotide sequence information represented by SEQ ID NOS.2 to 3431;
- (ii) classifying the isozyme identified in (i) into an isozyme having the same activity;
- (iii) mutating all genes encoding the isozyme having the same activity simultaneously; and
- (iv) examining productivity by a fermentation method of the compound selected in (i) of the coryneform bacterium which have been transformed with the gene obtained in (iii).
59. A method for breeding a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:2 to 3431, comprising the following:
- (i) arranging a function information of an open reading frame (ORF) represented by SEQ ID NOS:2 to 3431;
- (ii) allowing the arranged ORF to correspond to an enzyme on a known biosynthesis or signal transmission pathway;
- (iii) explicating an unknown biosynthesis pathway or signal transmission pathway of a coryneform bacterium in combination with information relating known biosynthesis pathway or signal transmission pathway of a coryneform bacterium;
- (iv) comparing the pathway explicated in (iii) with a biosynthesis pathway of a target useful product; and
- (v) transgenetically varying a coryneform bacterium based on the nucleotide sequence information to either strengthen a pathway which is judged to be important in the biosynthesis of the target useful product in (iv) or weaken a pathway which is Judged not to be important in the biosynthesis of the target useful product in (iv).
60. A coryneform bacterium, bred by the method of any one of claims 52 to 59.
61. The coryneform bacterium according to claim 60, which is a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium, or the genus tricrobacteritm.
62. The coryneform bacterium according to claim 61, wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
63. A method for producing at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid and an analogue thereof, comprising:
- culturing a coryneform bacterium of any one of claims 60 to 62 in a medium to produce and accumulate at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof;
- recovering the compound from the culture.
64. The method according to claim 63, wherein the compound is L-lysine.
65. A method for identifying a protein relating to useful mutation based on proteome analysis, comprising the following:
- (i) preparing
- a protein derived from a bacterium of a production strain of a coryneform bacterium which has been subjected to mutation breeding by a fermentation process so as to produce at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof, and
- a protein derived from a bacterium of a parent strain of the production strain;
- (ii) separating the proteins prepared in (i) by two dimensional electrophoresis;
- (iii) detecting the separated proteins, and comparing an expression amount of the protein derived from the production strain with that derived from the parent strain;
- (iv) treating the protein showing different expression amounts as a result of the comparison with a peptidase to extract peptide fragments;
- (v) analyzing amino acid sequences of the peptide fragments obtained in (iv); and
- (vi) comparing the amino acid sequences obtained in (v) with the amino acid sequence represented by SEQ ID NOS:3502 to 7001 to identifying the protein having the amino acid sequences.
66. The method according to claim 65, wherein the coryneform bacterium is a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium, or the genus bicrobacteium.
67. The method according to claim 66, wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
68. A biologically pure culture of Corynebacterium glutamicum AHP-3 (FERM BP-7382).
Type: Application
Filed: Dec 18, 2000
Publication Date: Dec 26, 2002
Inventors: Satoshi Nakagawa (Tokyo), Hiroshi Mizoguchi (Tokyo), Seiko Ando (Tokyo), Mikiro Hayashi (Tokyo), Keiko Ochiai (Tokyo), Haruhiko Yokoi (Tokyo), Naoko Tateishi (Tokyo), Akihiro Senoh (Tokyo), Masato Ikeda (Tokyo), Akio Ozaki (Hofu-shi)
Application Number: 09738626
International Classification: C12Q001/68; C12P019/34; C12M001/34;
