Abstract: The present invention provides a novel genetically modified Escherichia coli JM109 bearing accession number PTA 1579, containing the gene coding for poly-beta-hydroxybutyrate synthesis and a method of using this bacterium to produce poly-beta-hydroxybutyrate to the extent of 60% or more of the cell weight.

Abstract: This invention provides improved electroporation methods for transferring nucleic acids of interest into host cells, wherein the host cells are (1) suspended in a substantially non-ionic solution comprising at least one sugar or sugar derivative, (2) mixed with the nucleic acids of interest, and (3) electrically treated. Also, this invention provides for kits used in the method for transferring nucleic acids into host cells.

Abstract: This invention provides methods for producing and selecting host cells that better survive transformation treatment by subjecting host cells to conditions that alter them, subjecting the altered cells to transformation conditions, and selecting host cells that survive the transformation conditions. This invention also provides methods for transferring nucleic acids of interest into host cells, using cells that are better able to survive transformation treatment. Also, this invention provides kits for producing or selecting host cells in transformation treatments, as well as, kits comprising various host cells that may be utilized in transformation experiments.

Abstract: A microorganism is described which is transformed with DNAs which encode a hydantoinnase, a racemase, and a carbamoylase. As a result, the microorganism is able to degrade hydantoins directly to amino acids. A process for the production of the microorganism and a process for producing amino acids with the microorganism is also described.

Abstract: The present invention provides novel rapidly growing microorganisms and methods for their use in cloning or subcloning nucleic acid molecules. The rapid growing microorganisms of the present invention form colonies more rapidly than microorganisms typically used in molecular biology and thus provide a significant improvement in in vitro cloning methods used extensively in molecular biology. The invention also relates to kits and compositions used in the methods of the invention.

Abstract: The invention relates to the nucleotide sequence from sugarcane ubi9 polyubiquitin gene promoter, which is capable of directing constitutive expression of a nucleic acid sequence of interest that is operably linked to it. The sugarcane ubi9 promoter is useful in regulating expression of a nucleic acid sequence of interest in monocotyledonous and dicotyledonous plants.

Abstract: The present invention relates to a new plasmid originated from (Bifidobacterium) a recombinant expression vector and transformation method using the same. More particularly, the present invention relates to a plasmid pMG1 having nucleotide sequence represented by SEQ.ID.NO. 1; (Bifidobacterium longum) MG1 including the plasmid pMG1; and a shuttle vector which can be replicated in both (Bifidobacterium) and (E. coli), and comprises (Mob) gene having nucleotide sequence represented by SEQ.ID.NO. 2. (Rep) gene having nucleotide sequence represented by SEQ.ID.NO. 3 and a selection marker. The shuttle vector and the promoter of the present invention can be used for expressing target gene without additional purification process. The protein expressed from the target gene in (Bifidobacterium) can be added to food, therefore, the protein can be used for preparing food additives or oral vaccine.

Abstract: The present invention relates to methods and compositions which enable the propagation of vectors containing cDNAs whose presence has hitherto been toxic to conventional bacterial strains. It is based, at least in part, on the discovery that a bacterial strain having an insertional mutation in the malT gene of Escherichia coli tolerated the propagation of a mec-4 cDNA-containing plasmid which was toxic to other bacterial strains. The methods and compositions of the invention may be particularly useful in the propagation of cDNAs encoding membrane proteins. The present invention also provides for ion channel assay systems comprising MEC-2, MEC-4, MEC-10 or variants thereof.

Abstract: The present invention relates to a vector comprising sequences that permit direct transfer of the vector from one prokaryotic cell to another, such as by intergeneric conjugation. The invention also relates to methods of making and using the vector.

Abstract: A method for site-directed mutagenesis which achieves mutant strand selection by introducing two flanking mutagenic primers.

Abstract: The present invention is directed to an in vivo method of transferring DNA from a donor cell to a recipient cell. In a specific embodiment, the method comprises a highly efficient process of bacterial mating. In a preferred embodiment, selection for a recombinant plasmid against a parent host plasmid and donor plasmid is based on recircularization of the host plasmid by its recombination with a gene of interest such that it is now no longer cleaved by a restriction enzyme expressed in the recipient cell.

Abstract: In one aspect the invention provides methods for moving an insert nucleic acid molecule between vectors using site-specific recombination in vivo. In another aspect, the invention provides methods for the functional analysis of a genome using site-specific recombination in vivo. Another aspect of the invention provides methods for deleting a target genomic region by intra-molecular site-specific recombination. Further aspects provide vectors and kits for use in the methods of the invention.

Abstract: The present invention concerns compositions, apparatus and methods of use of recognition complexes, comprising biological sensors operably linked to an organic semiconductor. Multiple recognition complexes can be associated into a recognition complex system. The recognition complex system is of use to identify analytes, to separate biological sensors that bind to a target analyte from those that do not, to separate analytes that bind to a specific biological sensor from those that do not, and to prepare biological sensors with a high affinity for a particular analyte. The recognition complex system may be attached to a variety of surfaces, such as a chip, a flow cell, magnetic beads or non-magnetic beads. The biological sensor may be used for screening of, for example, a phage library, combinatorial chemistry library, plant tissue extract or animal tissue extract for inhibitors, activators or binding factors of bioactive molecules.

Abstract: This invention relates to a method for the production of D-(−)-3-hydroxybutyric acid, comprising the step of culturing a recombinant strain containing genes phbA, phbB, ptb and buk by fermentation. Preferably, the recombinant strain is a strain of E. coli. The method of the invention is simple, avoiding the technique of degrading polymer to produce D-(−)-3-hydroxybutyric acid. The present method also provides improved efficiency, lowers the complicated requirement for facilities as used in traditional chemical synthesis, simplifies the complicated technique flow, and omits the complicated chiral separation step. Therefore, the present method greatly reduces the costs associated with D-(−)-3-hydroxybutyric acid production. Also, with this invention, the problems such as environmental pollution of chemical synthesis and chiral separation are overcome.

Abstract: To provide a promoter, a recombinant DNA, a gene expression vector, an expression vector, and a transformant, which are capable of expressing a gene without inducing gene expression with an inducer; and a method for producing a protein and a kit therefor, which can be operated easily and performed inexpensively by convenient and inexpensive steps.

Abstract: The present invention relates to a modified phage display method for detecting and identifying target and target binders. The modified methods involve transforming host cells with two separate phages, one comprising a target or target library and the other comprising a target-binder library, and selecting to eliminate the non-paired targets and binders. Also related are complexes comprising target and target binders, as well as antibodies that bind to these complexes. Additionally, the present invention also relates to nucleic acids encoding target and target binders peptides. The invention also relates to the use of the sequence information inherent in the targets and target binders for target validation using in silico approaches. In addition, the invention also relates to diagnostics and therapeutics employing the disclosed target and target binder peptides or polynucletides and their use in small molecule drug discovery.

Abstract: Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).

Abstract: In an expression vector having a ColE1 replication system, the homology of the RNAI and RNAII of the ColE1 origin of replication to uncharged tRNAs is modified mutations in the coding region of the RNAI gene and corresponding mutations in the RNAII gene. The mutation results in one or more base exchanges in loop 1 and/or loop 2 and/or loop 3 of RNAI and RNAII. In methods using this vector for producing recombinant proteins, plasmid copy number is stably maintained. In methods for plasmid production, high plasmid copy numbers can be obtained.

Abstract: The invention relates to recombinant plasmid pSP1 harboring the gene vapk, which encodes alkalic protease VapK, and par gene, which is associated with the stability of plasmid, a microorganism Vibrio metschnikovii transformed therewith, and method for producing an alkaline protease VapK using the same microorganism.

Abstract: The present invention provides a recombinant plasmid vector comprising a kanamycin resistance gene, a promoter, an endoxylanase signal sequence, a nucleotide sequence coding for an oligopeptide consisting of 13 amino acids including 6 consecutive histidine residues, and a human granulocyte colony stimulating factor (hG-CSF) gene; an E. coli transformed with the said vector; and, a process for producing complete hG-CSF protein with high purity from the protein pool secreted by the said microorganism.

Abstract: The present invention provides a bacterium having a genome that is genetically engineered to be at least 2 to 14% smaller than the genome of its native parent strain. A bacterium with a smaller genome can produce a commercial product more efficiently. The present invention also provides methods for deleting genes and other DNA sequences from a bacterial genome. The methods provide precise deletions and seldom introduces mutations to the genomic DNA sequences around the deletion sites. Thus, the methods can be used to generate a series of deletions in a bacterium without increasing the possibility of undesired homologous recombination within the genome. In addition, some of the methods provided by the present invention can also be used for replacing a region of a bacterial genome with a desired DNA sequence.

Abstract: A fast method of transforming competent cells is described. The competent cells are thawed at room temperature or in a water bath. Plasmid DNAs and competent cells are mixed together, then the mixture is subject to heat shock treatment. After plating the mixture on a low-temperature selective medium by a low-temperature plating tool, the competent cells are cultured on the selective medium.

Abstract: A process for the preparation of D-pantothenic acid and/or salts thereof or feedstuffs additives comprising these by fermentation of microorganisms of the Enterobacteriaceae family, in particular those which already produce D-pantothenic acid, in which the nucleotide sequence(s) in the microorganisms which code(s) for the pepB gene is/are enhanced, in particular over-expressed.

Abstract: The invention is based on the discovery that recombinagenic oligonucleobases are active in prokaryotic cells that contain a strand transfer activity (RecA) and mismatch repair activity (MutS). Using this system a type of Duplex Mutational Vector termed a Heteroduplex Mutational Vector, was shown to be more active in prokaryotic cells than the types of mutational vectors heretofore tested. Further improvements in activity were obtained by replacing the tetrathymidine linker by a nuclease resistant oligonucleotide, such as tetra-2′-O-methyl-uridine, to link the two strands of the recombinagenic oligonucleobase and removing the DNA-containing intervening segment. The claims concern Duplex Mutational Vectors that contain the above improvements. In an alternative embodiment the claims concern the use of Duplex Mutational Vectors in prokaryotic cells.

Abstract: This invention provides improved electroporation methods for transferring nucleic acids of interest into host cells, wherein the host cells are (1) suspended in a substantially non-ionic solution comprising at least one sugar or sugar derivative, (2) mixed with the nucleic acids of interest, and (3) electrically treated. Also, this invention provides for kits used in the method for transferring nucleic acids into host cells.

Abstract: The present invention relates to the field of biotechnology. The invention provides a novel approach using tobacco mosaic virus omega leader sequence to enhance the solubility of the recombinant products in E. coli and the method of use therefore. The invention provides the utilization of tobacco mosaic virus omega leader sequence into E. coli expression vector, and the tobacco mosaic virus omega leader sequence containing expression vector can be used in combination with other available means to obtain higher expression or better solubility. The invention can be applied to biotechnological pharmaceutical industry, genetic engineering, biochemistry and molecular biology etc. The invention provides an expression vector pTORG, which is a highly efficient GST fusion expression vector, and can significantly enhance the yield of biologically active recombinant products.

Abstract: The present invention provides methods of preparing gene targeted mammalian cells having a targeted gene mutation methods of making gene targeted mice, and gene targeting vectors that are useful in these methods.

Abstract: The invention belongs to the field of thrombolysis and of tissue plasminogen activator (tPA) derivative production in prokaryotic cells. The invention relates to methods for the production of a recombinant DNA-derived tPA, a variant therof or a (Kringle 2 Serine) K2S molecule or a variant therof in prokaryotic cells, wherein the tPA or K2S or variant is secreted extracellularly as an active and correctly folded protein, and the prokaryotic cell contains and expresses a vector comprising the DNA coding for the tPA or K2S or variant operably linked to the DNA coding for the signal peptide OmpA. The invention further relates to specific K2S derivatives obtainable by the method. The invention further relates to the DNA molecules and the use of the DNA molecules in the methods.

Abstract: A temperature-sensitive plasmid which is capable of autonomous replication in Escherichia coli K-12 at 10-30° C., but, at a temperature of 33° C. or more, is incapable of autonomous replication in Escherichia coli K-12 or is distributed unhomogeneously upon the cell division of Escherichia coli K-12, thereby not to be stably carried within cells of Escherichia coli K-12 under said temperature, and which is incapable of autonomous replication in a microorganism belonging to the genus Escherichia other than Escherichia coli K-12 or is distributed unhomogeneously upon cell division of said microorganism at any temperature, thereby not to be stably carried within cells of said microorganism.

Abstract: The invention refers to a novel method for cloning DNA molecules using a homologous recombination mechanism between at least two DNA molecules comprising: a) providing a host cell capable of performing homologous recombination, b) contacting in said host cell a first DNA molecule which is capable of being replicated in said host cell with a second DNA molecule comprising at least two regions of sequence homology to regions on the first DNA molecule, under conditions which favour homologous recombination between said first and second DNA molecules and c) selecting a host cell in which homologous recombination between said first and second DNA molecules has occurred.

Abstract: The invention relates to advantageous processes for preparing heterologous proteins in prokaryotic host cells by improved codon use and/or expression of tRNAs which code for codons occurring rarely in said host cell.

Abstract: The present invention is directed to methods for producing gene targeting constructs by homologous recombination using mouse genomic libraries arrayed in yeast shuttle vectors. The invention is also directed to methods of using targeting constructs made by the methods to generate transgenic animals.

Abstract: Mismatch Repair Detection (MRD), a novel method for DNA-variation detection, utilizes bacteria to detect mismatches by a change in expression of a marker gene. DNA fragments to be screened for variation are cloned into two MRD plasmids, and bacteria are transformed with heteroduplexes of these constructs. Resulting colonies express the marker gene in the absence of a mismatch, and-lack expression in the presence of a mismatch. MRD is capable of detecting a single mismatch within 10 kb of DNA. In addition, MRD can analyze many fragments simultaneously, offering a powerful method for high-throughput genotyping and mutation detection.

Abstract: A method for in vivo production of a library in cells comprising a multitude of mutated genetic elements, wherein an error-prone polymerase is used in each ancestral cell to replicate all or a part of a genetic element independently of the host chromosomal replication machinery. The genetic element comprises i) an origin of replication from which replication is initiated, ii) optionally a genetic marker, e.g. a gene conferring resistance towards an antibiotic, iii) a gene encoding the polypeptide of interest. Also methods for the generation of a DNA sequence encoding a desired variant of a polypeptide of interest, and for the determination of such a DNA sequence are described.

Abstract: The gene encoding a 4-hydroxybutyryl-Co A transferase has been isolated from bacteria and integrated into the genome of bacteria also expressing a polyhydroxyalkanoate synthase, to yield an improved production process for 4HB-containing polyhydroxyalkanoates using transgenic organisms, including both bacteria and plants. The new pathways provide means for producing 4HB containing PHAs from cheap carbon sources such as sugars and fatty acids, in high yields, which are stable. Useful strains are obtaining by screening strains having integrated into their genomes a gene encoding a 4HB-CoA transferase and/or PHA synthase, for polymer production. Processes for polymer production use recombinant systems that can utilize cheap substrates. Systems are provided which can utilize amino acid degradation pathways, &agr;-ketoglutarate, or succinate as substrate.

Abstract: The complete nucleotide sequence of the genome of porcine adenovirus type 3 (PAV-3) is provided. Methods for construction of infectious PAV genomes by homologous recombination in procaryotic cells are provided. Recombinant PAV viruses are obtained by transfection of mammalian cells with recombinant PAV genomes. The PAV-3 genome can be used as a vector for the expression of heterologous nucleotide sequences, for example, for the preparation and administration of subunit vaccines to swine or other mammals. In addition, PAV-3 vectors can be used for gene therapy and expression of heterologous polypeptides. PAV-3 genome sequences can also be used for diagnostic purposes, to detect the presence of PAV-3 DNA in a subject or biological sample.

Abstract: The present invention provides a isolated bacteriophage useful as a tool for studying biological, biochemical, physiological and genetic properties of actinomycetes and other organisms which comprises a novel strain of Saccharomonospora having certain specified characteristics. The invention also relates to a process for the isolation of the said bacteriophage and/or DNA phage and to a novel universal growth medium which is particularly useful in the said process. Another embodiment of the process relates to a cloning vector which comprises a plasmid or bacteriophage comprising the phage DNA of the invention.

Abstract: This invention provides improved electroporation methods for transferring nucleic acids of interest into host cells, wherein the host cells are (1) suspended in a substantially non-ionic solution comprising at least one sugar or sugar derivative, (2) mixed with the nucleic acids of interest, and (3) electrically treated. Also, this invention provides for kits used in the method for transferring nucleic acids into host cells.

Abstract: The present invention provides the promoter clone discovery of a glucoamylase gene of a starch utilizing yeast strain Schwanniomyces castellii. The isolated glucoamylase promoter is an inducible promoter, which can regulate strong gene expression in starch culture medium.

Abstract: An objective of this invention is to provide an effective process for producing a recombinant protein using E. coli and a technique therefor.

Abstract: The present invention relates to a method for producing mutants of a fatty acid desaturase having a substantially increased activity towards fatty acid substrates with chains containing fewer than 18 carbons relative to an unmutagenized precursor desaturase having an 18 carbon atom chain length substrate specificity. The method involves inducing one or more mutations in the nucleic acid sequence encoding the precursor desaturase, transforming the mutated sequence into an unsaturated fatty acid auxotroph cell such as MH13 E. coli, culturing the cells in the absence of supplemental unsaturated fatty acids, thereby selecting for recipient cells which have received and which express a mutant fatty acid desaturase with an elevated specificity for fatty acid substrates having chain lengths of less than 18 carbon atoms. A variety of mutants having 16 or fewer carbon atom chain length substrate specificities are produced by this method.

Abstract: The present invention provides a novel genetically modified Escherichia coli JM109 bearing accession number PTA 1579, containing the gene coding for poly-beta-hydroxybutyrate synthesis and a method of using this bacterium to produce poly-beta-hydroxybutyrate to the extent of 60% or more of the cell weight.

Abstract: The disclosure below provides a protein export system for efficiently producing recombinant protein from a host cell. In a preferred embodiment, the protein export system utilizes protein export machinery endogenous to the host bacterium into which the protein export system vector is introduced.

Abstract: The present invention provides a method for transferring and expressing heterologous DNA in a non-plant host cell. The vector used in this method includes a backbone having a first origin of replication capable of maintaining heterologous DNA as a single copy in an Escherichia coli host cell. The vector further includes a unique restriction endonuclease cleavage site for insertion of heterologous DNA, and left and right Agrobacterium T-DNA border sequences flanking the unique restriction endonuclease cleavage site. In certain host cells, the T-DNA border sequences allow introduction of heterologous DNA located between the left and right T-DNA border sequences into a host cell. In preferred embodiments, the vector includes a second origin of replication capable of maintaining heterologous DNA as a single copy in a host cell such as Agrobacterium species or other prokaryotic cells.

Abstract: The present invention relates to the fields of microbiology and microbial genetics. More specifically, the invention relates to novel bacterial strains and processes employing these strains for the fermentative production of amino acids such as threonine.

Abstract: Disclosed herein are novel phenol oxidizing enzymes naturally-produced by strains of the species Stachybotrys which possess a pH optima in the alkaline range and which are useful in modifying the color associated with dyes and colored compounds, as well as in anti-dye transfer applications. Also disclosed herein are biologically-pure cultures of strains of the genus Stachybotrys, designated herein Stachybotrys parvispora MUCL 38996 and Stachybotrys chartarum MUCL 38898, which are capable of naturally-producing the novel phenol oxidizing enzymes. Disclosed herein is the amino acid and nucleic acid sequence for Stachybotrys phenol oxidizing enzymes as well as expression vectors and host cells comprising the nucleic acid. Disclosed herein are methods for producing the phenol oxidizing enzyme as well as methods for constructing expression hosts.

Abstract: A method for producing a polypeptide product which is substantially free of an undesired protein, the process comprising culturing a host cell which is able to express said polypeptide product and which is able to express said undesired protein only in a mutant form which form has the activity of the corresponding native protein under culture conditions but is unstable under conditions at which the said polypeptide product remains stable; and recovering the desired product, wherein either the host cell culture or the recovered product is subjected for a sufficient period of time to conditions under which the undesired protein is unstable so as to denature the undesired protein The method allows the production of products which are substantially free of particular undesired contaminants.

Abstract: The present invention provides methods for directing the evolution of microorganisms comprising the use of mutator genes and growth under conditions of selective pressure. The method discloses mutator genes which can be used in the methods of the present invention and provides ATCC deposits which exemplify the evolved microorganisms produced by the methods.

Abstract: Methods are disclosed for identifying an RNA fragment that mimics the structure of a defined or undefined target RNA molecule to which a compound binds inside of a cell resulting in retardation of cell growth or cell death. Methods using these RNA fragments for identifying unknown compounds of pharmaceutical interest, and for identifying unknown RNA targets for use in treating disease are disclosed. These methods and compositions are used in screening for novel antibiotics, bacteriostatics, or modifications thereof or for identifying compounds useful to alter expression levels of proteins encoded by mRNA. The methods involve providing random DNA fragments from DNA which encodes RNA target molecules, cloning such fragments to create a plasmid library of same; transfecting cells which contain the native RNA target molecule with the plasmid library and exposing the cells to one or more of test compounds.

Abstract: This invention provides improved electroporation methods for transferring nucleic acids of interest into host cells, wherein the host cells are (1) suspended in a substantially non-ionic solution comprising at least one sugar or sugar derivative, (2) mixed with the nucleic acids of interest, and (3) electrically treated. Also, this invention provides for kits used in the method for transferring nucleic acids into host cells.