Abstract: The present invention relates to a system for regulating gene expression in prokaryotes using modified tetracycline repressor proteins. In particular, the present invention relates to modified tetracycline repressor proteins that exhibit a “reverse” phenotype in prokaryotic organisms, nucleic acids encoding these repressor proteins, methods for identifying and preparing these proteins, and methods for using these proteins for regulating gene expression in prokaryotic organisms, in drug screening assays and for identifying non-antibiotic compounds that are specific inducers of these modified repressor proteins.

Abstract: The present invention is directed to variants of antigens comprising folate binding protein epitopes as a composition associated with providing immunity against a tumor in an individual. The variant is effective in inducing cytotoxic T-lymphocytes but preferably not to the extent that they become sensitive to silencing by elimination, such as by apoptosis, or by anergy, as in unresponsiveness.

Abstract: A process for purification of recombinant porphobilinogen deaminase (rhPBGD) on an industrial scale by starting from a rhPBGD containing extract obtained from a fermentation of a recombinant cell capable of expressing the rhPBGD and the use of the purified product for the preparation of a medicament.

Abstract: Regulation of expression of programmed cell death, including senescence, in plants is achieved by integration of a gene or gene fragment encoding senescence-induced deoxyhypusine synthase, senescence-induced eIF-5A or both into the plant genome in antisense orientation. Plant genes encoding senescence-induced deoxyhypusine synthase and senescence-induced eIF-5A are identified and the nucleotide sequences of each, alone and in combination are used to modify senescence in transgenic plants.

Abstract: Isolated nucleic acid sequences and polypeptides encoded thereby for epothilone B hydroxylase and mutants and variants thereof and a ferredoxin located downstream from the epothilone B hydroxylase gene are provided. Also provided are vectors and cells containing these vectors. In addition, methods for producing recombinant microorganisms, methods for using these recombinant microorganism to produce hydroxyalkyl-bearing epothilones and an epothilone analog produced by a mutant of epothilone B hydroxylase are provided.

Abstract: The present invention describes a novel recombinant NADH recycling system that is used as a process for producing reduced compounds. In a specific embodiment, the reduced compounds include ethanol, succinate, lactate, a vitamin, a pharmaceutical and a biodegraded organic molecule. The NADH recycling system effects metabolic flux of reductive pathways in aerobic and anaerobic environments.

Abstract: This invention relates to an isolated nucleic acid fragment encoding a tetrahydrofolate metabolic enzyme. The invention also relates to the construction of a chimeric gene encoding all or a portion of the tetrahydrofolate metabolic enzyme, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the tetrahydrofolate metabolic enzyme in a transformed host cell.

Abstract: The invention provides DNA coding for cytochrome P450 monooxygenases of the CYP79 family catalyzing the conversion of an aliphatic or aromatic amino acid or chain-elongated methionine homologue to the corresponding oxime. Preferred embodiments of the invention are enzymes catalyzing the conversion of L-Valine and L-Isoleucine such as the cassava enzymes CYP79D1 and CYP79D2, enzymes catalyzing the conversion of tyrosine such as the Triglochin maritima enzymes CYP79E1 and CYP79E2, enzymes catalyzing the conversion of tryptophan to the corresponding oxime indole-3-acetaldoxime such as the Arabidopsis thaliana enzyme CYP79A2 and the Brassica napus enzyme CYP79B5, and enzymes catalyzing the conversion of a chain-elongated methionine homologue such as the Arabidopsis thaliana enzymes CYP79F1 and CYP79F2. Transgenic expression of said DNA or parts thereof in plants can be used to manipulate the biosynthesis of corresponding glucosinolates or cyanogenic glucosides.

Abstract: The invention provides active, non-naturally occurring mutants of beetle luciferases and DNAs which encode such mutants. A mutant luciferase of the invention differs from the corresponding wild-type luciferase by producing bioluminescence with a wavelength of peak intensity that differs by at least 1 nm from the wavelength of peak intensity of the bioluminescence produced by the wild-type enzyme. The mutant luciferases and DNAs of the invention are employed in various biosensing applications.

Abstract: Compositions and methods for use in generating and selecting genetically modified cells are provided. The compositions include selectable markers and selection systems based thereon. Also provided are methods for the introduction and expression of heterologous nucleic acids in host animals that use the compositions and methods for generating and selecting genetically modified cells.

Abstract: This invention relates to an isolated nucleic acid fragment encoding a histidine biosynthetic enzyme. The invention also relates to the construction of a chimeric gene encoding all or a portion of the histidine biosynthetic enzyme, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the histidine biosynthetic enzyme in a transformed host cell.

Abstract: A process is disclosed for the preparation of a recombinantly expressed fusion product comprised of a proteinaceous tag and a soluble protein of interest and the separation of the fusion product from the host cell in which it is expressed. The tag and in turn the fusion product is insoluble in the host cell lysate solution and the fusion product is separated therefrom by centrifugation or filtration.

Abstract: The invention relates to compositions comprising a first fusion protein comprising a first polypeptide domain and a R. reniformis luciferase and a second fusion protein comprising a second polypeptide domain and a R. reniformis GFP. The invention also relates to compositions comprising one or more polynucleotides encoding a first fusion protein comprising a first polypeptide domain and a R. reniformis luciferase and a second fusion protein comprising a second polypeptide domain and a R. reniformis GFP. The invention also relates to methods and kits for detecting protein-protein interactions, determining the location of a protein-protein interaction, identifying cells wherein there is a protein-protein interaction of interest, and screening for a candidate modulator that increases or decreases the amount of a protein-protein interaction.

Abstract: The present invention has an object of providing a novel fructosyl peptide oxidase having superior physicochemical properties such as stability that is useful as an enzyme for clinical diagnosis, and an object of providing a method for producing the fructosyl peptide oxidase.

Abstract: The present invention relates to polynucleotides that encode proteins having OpcA enzymatic activity. These polynucleotides can be used for increasing lysine biosynthesis in Coryneform glutamicum.

Abstract: The invention provides a biosynthetic gene cluster for mitomycin, as well as methods of using gene(s) within the cluster to alter biosynthesis.

Abstract: Transgenic microbial strains are provided which contain the genes required for PHA formation integrated on the chromosome. The strains are advantageous in PHA production processes, because (1) no plasmids need to be maintained, generally obviating the required use of antibiotics or other stabilizing pressures, and (2) no plasmid loss occurs, thereby stabilizing the number of gene copies per cell throughout the fermentation process, resulting in homogeneous PHA product formation throughout the production process. Genes are integrated using standard techniques, preferably transposon mutagenesis. In a preferred embodiment wherein mutiple genes are incorporated, these are incorporated as an operon. Sequences are used to stabilize mRNA, to induce expression as a function of culture conditions (such as phosphate concentration), temperature, and stress, and to aid in selection, through the incorporation of selection markers such as markers conferring antibiotic resistance.

Abstract: The present invention relates to an enzymatic composition capable of killing or inhibiting microbial cells or micro-organisms, e.g. in laundry, on hard surfaces, in water systems, on skin, on teeth or on mucous membranes. The present invention also relates to the use of said enzymatic composition for preserving food products, cosmetics, paints, coatings, etc.

Abstract: The present invention provides an isolated, purified and characterized human tyrosine hydroxylase (hTH) promoter nucleic acid sequence. The invention further provides a method of selecting TH positive (TH+) cells by preparing a construct comprising a hTH promoter operably linked to a heterologous nucleic acid sequence, for example, green fluorescent protein encoding sequence, and transfecting cells, particularly stem cells, with the construct. The invention also provides a hTH promoter, useful in gene therapeutic applications in driving therapeutic genes or other nucleic acid sequences operably linked to the hTH promoter. Additionally, the invention provides cell lines and transgenic animals expressing a transgene comprising the hTH promoter operably linked to a heterologous sequence, which cell lines and transgenic animals are useful for isolating TH+ cells for transplantation or for screening of therapeutic agents that affect TH+ function.

Abstract: Enzymatic process for the preparation of cephalosporanic 7?-(4-carboxybutananude) acid by using the modified enzyme D-aminoacid oxidase of Trigonopsis variabilis produced in Escherichia coli. The process for the expression of the enzyme comprises: (I) isolating the DNA corresponding to gene which codes for the enzyme D-aminoacid oxidase; (II) removing the intron which is contained in said gene; (III) inserting the DNA fragment obtained into the plasmide which is capable of replication in Escherichia coli; (IV) fusing at the extremity 5′ of the structural region of the gene a synthetic assembler which contains a nucleotide sequence which codes for six histidines; (V) transforming a strain of Escherichia coli with the resulting recombinant plasmide; (VI) cultivating the transformed cells of Escherichia coli; and (VII) recovering the enzyme D-aminoacid oxidase of the former cultivation operation through affinity chromatography.

Abstract: The present invention provides a vector encoding a detectable cell surface marker and a suicide construct, and cells and a non-human mammal transduced with this vector. Introduction of lymphocytes transduced with this vector, after allogeneic bone marrow transplantation, serves to treat or prevent complications from the bone marrow transplant, including graft versus host disease.

Abstract: Methods of reducing cystine containing animal and plant proteins, and improving dough and baked goods' characteristics is provided which includes the steps of mixing dough ingredients with a thiol redox protein to form a dough and baking the dough to form a baked good. The method of the present invention preferably uses reduced thioredoxin with wheat flour which imparts a stronger dough and higher loaf volumes. Methods for reducing snake, bee and scorpion toxin proteins with a thiol redox (SH) agent and thereby inactivating the protein or detoxifying the protein in an individual are also provided. Protease inhibitors, including the Kunitz and Bowman-Birk trypsin inhibitors of soybean, were also reduced by the NADP/thioredoxin system (NADPH, thioredoxin, and NADP-thioredoxin reductase) from either E. coli or wheat germ. When reduced by thioredoxin, the Kunitz and Bowman-Birk soybean trypsin inhibitors lose their ability to inhibit trypsin.

Abstract: Regulation of expression of programmed cell death, including senescence, in plants is achieved by integration of a gene or, gene fragment encoding senescence-induced deoxyhypusine synthase, senescence-induced eIF-5A or both into the plant genome in antisense orientation. Plant genes encoding senescence-induced deoxyhypusine synthase and senescence-induced eIF-5A are identified and the nucleotide sequences of each, alone and in combination are used to modify senescence in transgenic plants.

Abstract: Regulation of expression of programmed cell death, including senescence, in plants is achieved by integration of a gene or gene fragment encoding senescence-induced deoxyhypusine synthase, senescence-induced eIF-5A or both into the plant genome in antisense orientation. Plant genes encoding senescence-induced deoxyhypusine synthase and senescence-induced eIF-5A are identified and the nucleotide sequences of each, alone and in combination are used to modify senescence in transgenic plants.

Abstract: A naturally occurring or recombinant urate oxidase (uricase) covalently coupled to poly(ethylene glycol) or poly(ethylene oxide) (both referred to as PEG), wherein an average of 2 to 10 strands of PEG are conjugated to each uricase subunit and the PEG has an average molecular weight between about 5 kDa and 100 kDa. The resulting PEG-uricase conjugates are substantially non-immunogenic and retain at least 75% of the uricolytic activity of the unmodified enzyme.

Abstract: Novel DNA constructs are provided which may be used as molecular probes or inserted into a plant host to provide for modification of transcription of a DNA sequence of interest in cotton fiber, particularly in very early fiber development. The DNA constructs comprise a cotton fiber transcriptional initiation regulatory region associated with a gene which is expressed in cotton fiber.

Abstract: This invention identifies a novel family of bilin reductases. Designated herein HY bilin reductases, the enzymes of this invention are useful in a wide variety of contexts including but not limited to the conversion of biliverdins to phytobilins and the assembly of holophytochromes or phytofluors.

Abstract: The present invention is directed to a new NADH oxidase from Lactobacillus, nucleic acids encoding the NADH oxidase, methods of producing the NADH oxidase, as well as there use in producing improved NADH oxidase enzymes and for producing chrial enantiomer-enriched organic compounds, such as alcohols and/or amino acids.

Abstract: Novel genes encoding homologous immunoreactive thio-disulfide oxidoreductases, or disulfide bond formation (Dsb) proteins from Ehrlichia chaffeensis and Ehrlichia canis are disclosed. While the E. chaffeensis and E. canis Dsb proteins are at most only 31% or less homologous to other known Dsb proteins, the Ehrlichia Dsbs contain a cysteine active site, Cys-Gly-Tyr-Cys, similar to those in known Dsb proteins. As predicted by 15-amino acid identical N-terminal signal peptides, the proteins are primarily localized in the periplasm of E. chaffeensis and E. canis, possibly playing a role in antigenicity and pathogenesis. The present invention provides the nucleotide and amino acid sequences and expression vectors for the E. chaffeensis and E. canis dsb genes, antisera directed against the proteins, and kits to determine whether an individual or animal is infected with a given species of Ehrlichia.

Abstract: An isolated gene encodes for a L-proline-3-hydroxylase having the amino acid sequence of SEQ ID NOS: 1, 2, 15, 16 and 17, or for a protein having enzymatic activity to hydroxylate the 3-position of L-proline and to act on free L-proline in the presence of 2-ketoglutaric acid and divalent iron ions to produce cis-3-hydroxy-L-proline. The gene is inserted into a vector and the resulting recombinant DNA is used to create a transformant. L-proline-3-hydroxylase is produced by culturing the transformant in a medium.

Abstract: The invention provides a human lysyl hydroxylase-like protein (HLHLP) and polynucleotides which identify and encode HLHLP. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for treating or preventing disorders associated with expression of HLHLP.

Abstract: The invention features a novel gene encoding methionine synthase reductase. The invention also features a method for detecting an increased likelihood of hyperhomocysteinemia and, in turn, an increased or decreased likelihood of neural tube defects, cardiovascular disease, or cancer. The invention also features therapeutic methods for treating and/or reducing the risk of cardiovascular disease, cancer, or neural tube defects. Also provided are the sequences of the human methionine synthase reductase gene and protein and compounds and kits for performing the methods of the invention.

Abstract: A highly conserved active site helix present within the P-450 superfamily of proteins is found also in monoamine oxidase (MAO) B, a major enzyme that catalyzes deamination of neuro- and vaso-active amines in the nervous system of mammals. Mutation within the conserved region of the MAO B enzyme directly reduces MAO B's activity and alters its pH profile, which allows for indirect regulation of the cellular neurotransmitters and vasoamines.

Abstract: Genes encoding phenylalanine ammonia-lyase (PAL), tyrosine ammonia lyase (TAL) and phenylalanine hydroxylase (PAH) have been introduced into a host organism for the production of Para-hydroxycinnamic acid (PHCA). The introduction of these genes results in the redirection of carbon flow in the host, optimizing the flow of carbon from glucose to PHCA. The intermediates, tyrosine and cinnamic acid are also produced.

Abstract: Genes and proteins involved in the biosynthesis of benzodiazepines by microorganisms, including the genes and proteins forming the biosynthetic loci for the benzodiazepine anthramycin from Streptomyces refuineus subsp. thermotolerans. The genes and proteins allow direct manipulation of benzodiazepines and related chemical structures via chemical engineering of the enzymes involved in the biosynthesis of anthramycin.

Abstract: The present invention relates to methods for stimulating root growth and/or enhancing the formation of lateral or adventitious roots and/or altering root geotropism comprising expression of a plant cytokinin oxidase or comprising expression of another protein that reduces the level of active cytokinins in plants or plant parts.

Abstract: Plant nucleic acid compositions encoding polypeptide products with diacylglyceride acyltransferase (DGAT) activity, as well as the polypeptide products encoded thereby and methods for producing the same, are provided. Methods and compositions for modulating DGAT activity in a plant, particularly DGAT activity in plant seeds, and transgenic plants with altered DGAT activity are provided. Such plants and seeds are useful in the production of human food and animal feedstuff, and have several other industrial applications. Also provided are methods for making triglycerides and triglyceride compositions, as well as the compositions produced by these methods. The subject methods and compositions find use in a variety of different applications, including research, medicine, agriculture and industry.

Abstract: Disclosed is a family of P450 monooxygenases, each member of which regioselectively oxidizes avermectin to 4″-keto-avermectin. The P450 monooxgenases find use in methods and formulations for making emamectin from avermectin. Also disclosed are methods for purifying the P450 monooxygenases of the invention, binding agents that specifically bind to the P450 monooxygenases of the invention, and genetically engineered cells that express the P450 monooxygenases of the invention. Also disclosed are ferredoxins and ferredoxin reductases that are active with the P450 monooxygenases of the invention.

Abstract: Luciferase enzymes with greatly increased thermostability, e.g., at least half lifes of 2 hours at 50° C., cDNAs encoding the novel luciferases, and hosts transformed to express the luciferases, are disclosed. Methods of producing the luciferases include recursive mutagenesis. The luciferases are used in conventional methods, some employing kits.

Abstract: There is provided novel indole oxidase activity isolated from P. putida, such indole oxidase activity is believed to be useful in the biosynthetic production of indigo from its precursor indole. Also provided are compositions of matter comprising the indole oxidase and methods for producing such.

Abstract: The sequencing and analysis of the complete nucleotide sequence of symbiotic plasmid pNGR234a isolated from Rhizobium sp. NGR234. The complete sequence of pNGR234a is presented. The analysis includes the identification of a number of novel ORFs and the proteins expressible therefrom which have been ascribed putative functions.

Abstract: The present invention relates to new NOS variants or mutants which contain structural alterations in the site of Akt dependent phosphorylation. The altered NOS proteins or peptides, especially the human eNOS proteins or peptides, Akt proteins or polypeptides and their encoding nucleic acid molecules are useful as gene therapy agents for the treatment of diseases including post angioplasty restenosis, hypertension, atherosclerosis, heart failure, diabetes and diseases with defective angiogenesis. NOS proteins and peptides are also useful in methods of screening for agents which modulate NOS activity.

Abstract: The present invention provides amino alcohol dehydrogenases which catalyze a reaction to produce keto alcohols, ketones, aldehydes, keto acids from corresponding amino alcohols, amines, and amino acids in the presence of NAD+, and a reaction to produce amino alcohols, amines, amino acids from corresponding keto alcohols, ketones, aldehydes, and keto acids in the presence of NADH and ammonium ions. Also provided is a method for producing the enzymes and uses of the enzyme. The enzymes of the present invention can be obtained from microorganisms belonging to the genera Streptomyces, Pseudomonas, Burkholdenia, or Arthrobacter.

Abstract: DNA sequences are provided which encode the enzymes required for clavulanic acid synthesis. A process is provided for producing clavulanic acid in a transformant of a non-clavulanate-producing host.

Abstract: Recombinant lentiviral vectors having a region encoding a functional &bgr;-globin gene; and large portions of the &bgr;-globin locus control regions which include DNase I hypersensitive sites HS2, HS3 and HS4 provides expression of &bgr;-globin when introduced into a mammal, for example a human, in vivo. Optionally, the vector further includes a region encoding a dihydrofolate reductase. The vector may be used in treatment of hemoglobinopathies, including &bgr;-thalessemia and sickle-cell disease. For example, hematopoietic progenitor or stem cells may be transformed ex vivo and then restored to the patient. Selection processes may be used to increase the percentage of transformed cells in the returned population. For example, a selection marker which makes transformed cells more drug resistant than un-transformed cells allows selection by treatment of the cells with the corresponding drug.

Abstract: The present invention relates, in general, to a method of producing L-amino acids comprising culturing altered bacterial cells having increased amounts of NADPH as compared to unaltered bacterial cells whereby L-amino acids yields from said altered bacterial cells are greater than yields from unaltered bacterial cells. The invention also relates to a gene encoding phosphoglucoisomerase.

Abstract: A method for treating a porous article by efficiently performing macromolecularization in a porous article using an enzyme having a polyphenol oxidizing activity in an alkaline pH region, a phenolic compound and/or an aromatic amine compound, a composition for use in the treatment method, and treated products from the porous article obtained by the treatment method which are given or increased in strength, wear resistance, weatherability, rust-preventing properties, flame resistance, antibacterial properties, antiseptic properties, sterilizing properties, insect-repellent properties, insecticidal properties, antiviral properties, organism-repellent properties, adhesiveness, chemical agent-slow-releasing properties, coloring properties, dimension stability, crack resistance, deodorizing properties, deoxidizing properties, humidity controlling properties, moisture conditioning properties, water repellency, surface smoothness, bioaffinity, ion exchangeability, formaldehyde absorbing properties, chemical agent el

Abstract: This invention is directed to a DNA sequence comprising a nucleotide sequence encoding a variant paraoxonase protein and to said variant paraoxonase protein as well as a method and a kit for detecting a risk of cancer, coronary or cerebrovascular disease, hypertension, type 2 diabetes , dementia, joint arthrosis, cataract, or sensitivity to organophosphorus compounds in a subject, the method comprising isolating genomic DNA from said subject, determining the allelic pattern for the codon 102 of the paraoxonase encoding PON1 gene in the genomic DNA, identification of Ile101Val mutation indicating said risk being increased and for targeting paraoxonase activity modulating therapies. Further this invention relates to transgenic animals comprising a human DNA molecule encoding said variant paraoxonase and to a method of phenotype-targeted gene sequencing.

Abstract: This invention provides methods for practical enzymatic conversion of GDP-mannose to GDP-fucose. These methods are useful for efficient synthesis of reactants used in the synthesis of fucosylated oligosaccharides.

Abstract: The present invention makes use of a chimeric gene that, when incorporated into an appropriate host, results in the overproduction of S-adenosylmethionine without the need to supply the host with a source of untransformed methionine. The need for the methionine source is eliminated because the appropriately chosen host manufactures the amino acid on its own, and when the host is modified by the inclusion of a chimeric gene of the present invention, it transforms the methionine that is produced into S-adenosylmethionine. In addition, the same chimeric gene causes accumulation of methionine and increase in folate content in the same host. One form of the present invention is a fused gene encoding for methylene tetrahydrofolate reductase (MTHFR) made up of an N-terminal domain from a yeast organism and a C-terminal domain from a plant species. An example of a suitable plant species is Arabidopsis thaliana. An example of a suitable yeast organism is Saccharomyces cerevisiae.