Abstract: Disclosed herein are improved plasmid shuttle vectors, vaccines based on them, and methods related to their construction and use. Particular arrangements of functional elements of such plasmids, namely origins of replication and eukaryotic transcription/translation control elements, which give rise to generally undesirable side-products upon propagation of the plasmids in bacterial culture are disclosed. These side-products apparently arise as terminated replication intermediates. Strategies both to eliminate accumulation of these side-products, and to make them useful as a vaccine adjuvant, are described.
Abstract: A method for introducing and stabilizing heterologous and recombinant genes in a thermophilic host in which a characteristic gene defining a detectable host characteristic is inactivated or deleted from the thermophilic host, resulting in a modified thermophilic host expressing an absence of the detectable host characteristic. A DNA fragment of interest is inserted into the modified thermophilic host together with an intact characteristic gene, whereby the detectable host characteristic is restored to the thermophilic host, thereby enabling detection and confirmation of successful transformation using plasmid vectors and integration of the DNA fragment into the chromosome of the thermophilic host.
Type:
Application
Filed:
February 28, 2002
Publication date:
September 11, 2003
Inventors:
Kevin J. Kayser, Ho-Shin Park, John J. Kilbane
Abstract: The invention concerns a new tool for efficient mutagenesis enabling the generation of a collection of mutants in fungi by random insertion of a characterized Fusarium oxysporum Impala transposon in the genome of said fungi. The invention also concerns the resulting mutants.
Abstract: Disclosed and claimed are novel toxins and genes obtainable from Bacillus laterosporus isolates disclosed herein. In preferred embodiments, the subject genes and toxins are used to control Western corn rootworm.
Type:
Grant
Filed:
September 28, 2001
Date of Patent:
August 12, 2003
Assignee:
Mycogen Corporation
Inventors:
H. Ernest Schnepf, Kenneth E. Narva, Brian A. Stockhoff, Stacey Finstad Lee, Mikki Walz, Blake Sturgis
Abstract: A method for identifying a nucleic acid which codes for a polypeptide factor affecting the covalent bonding of polypeptides to the surface of Gram-positive bacteria, comprising the following steps:
Type:
Application
Filed:
July 23, 2002
Publication date:
July 31, 2003
Applicant:
Evotec BioSystems AG of Schnackenburgallee 114
Inventors:
Andreas Strauss, Gunther Thumm, Johannes Pohlner, Friedrich Gotz
Abstract: A process for the preparation of L-amino acids, in which the following steps are carried out,
a) fermenting the desired L-amino acid-producing bacteria in which at least the glyA gene is attenuated, in particular by removal of the natural promoter, and optionally
b) concentrating the desired product in the medium or in the cells of the bacteria and
c) isolating the L-amino acid,
and optionally bacteria in which further genes of the biosynthesis pathway of the desired L-amino acid are additionally amplified are employed, or bacteria in which the metabolic pathways which reduce the formation of the desired L-amino acid are at least partly eliminated are employed, and nucleotide sequences of the lacI-tac-5′glyA or lacI-tac-glyA unit.
Abstract: This invention relates to an isolated nucleic acid fragment encoding scorpion toxins that are K-channel modifiers. The invention also relates to the construction of a chimeric gene encoding all or a substantial portion of the K-channel modifier, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the K-channel modifier in a transformed host cell.
Type:
Grant
Filed:
June 1, 2001
Date of Patent:
July 15, 2003
Assignee:
E. I. du Pont de Nemours and Company
Inventors:
Rafael Herman, James F. Wong, Albert L. Lu, James K. Presnail, Jian-Ming Lee
Abstract: Provided is the active site of human gamma glutamyl hydrolase. The active site resides in amino acid residues 110, 171, 220 and 222 of SEQ ID NO:1. Thus provided is an inactive gamma glutamyl hydrolase protein, as well as a fragment thereof. A method of inactivating a gamma glutamyl hydrolase protein is also provided, as is a molecule capable of binding to one or more of amino acid residues 110, 171, 220 or 222 of SEQ ID NO:1 which can be used in such a method. A method for identifying a molecule that inactivates gamma glutamyl hydrolase is provided, as is a nucleic acid molecule encoding the inactive gamma glutamyl hydrolase.
Type:
Grant
Filed:
June 4, 1999
Date of Patent:
June 3, 2003
Assignee:
Health Research Incorporated
Inventors:
John H. Galivan, Thomas J. Ryan, Ivan E. Auger
Abstract: The present invention is directed to a process for the production of a peptide, polypeptide, or protein having a predetermined property. In accordance with one embodiment, the process begins by producing by way of synthetic polynucleotide coupling, stochastically generated polynucleotide sequences. A library of expression vectors containing such stochastically generated polynucleotide sequences is formed. Next, host cells containing the vectors are cultured so as to produce peptides, polypeptides, or proteins encoded by the stochastically generated polynucleotide sequences. Screening or selection is carried out on such host cells to identify a peptide, polypeptide, or protein produced by the host cells which has the predetermined property. The stochastically generated polynucleotide sequence which encodes the identified peptide, polypeptide, or protein is then isolated and used to produce the peptide, polypeptide, or protein having the predetermined property.
Abstract: The present invention relates to host cells suitable for expressing genes under the direction of foreign RNA polymerases and to providing very low levels of expression of such genes and RNA polymerases in the absence of induction.
Abstract: Nucleic acid molecules encoding an alternansucrase are provided. Moreover, vectors, host cells and plant cells transformed by the herein-described nucleic acid molecules and plants containing them are provided. Furthermore, methods are described for preparing transgenic plants which synthesize the carbohydrate alternan, because of the insertion of nucleic acid molecules encoding an alternansucrase. Moreover, methods for preparing alternan and products resulting from them are provided.
Type:
Grant
Filed:
February 8, 2000
Date of Patent:
May 27, 2003
Assignee:
Max-Planck-Gesellschaft zur Forderung der Wissenschaften
e.v.
Inventors:
Jens Kossmann, Thomas Welsh, Martin Quanz, Karola Knuth
Abstract: A screening method which applies the principle of synthetic lethality to a gene of interest in non-yeast eukaryotic cells. The method may be used to screen either a chemical library in order to identify a molecule having a gene-specific lethal property in the cells, or to screen a group of DNA molecules in order to identify among them one or more modulators of gene function which are synergistically lethal to the cells together with an incapacitated gene of interest. Also described are episomal survival plasmids and kits which may be used with the method.
Type:
Grant
Filed:
September 8, 1999
Date of Patent:
May 27, 2003
Assignee:
Ramot University Authority for Applied Research &
Industrial Development Ltd.
Inventors:
Dan Canaani, Yoram Oron, Arnold Simons, Naomi Dafni, Iris Dotan
Abstract: A method for detecting protein-protein interactions is provided, in which two fusion proteins are prepared and allowed to interact with each other in yeast cells. The interaction between the two fusion proteins leads to protein trans-splicing, generating an active and detectable reporter.
Abstract: The present invention relates to novel bacterial strains useful for the production of 2-keto-L-gulonic acid. The present invention further relates to the use of these strains for the production of 2-keto-L-gulonic acid by fermentative conversion of L-sorbose. The present invention further relates to the use of these novel bacterial strains for the production of pyrroloquinoline quinone and a nontoxic lipopolysaccharide. Also described is the strains of the present invention transformed by a vector.
Abstract: An isolated DNA having the promoter sequence of the hex gene of P. chrysogenum or a DNA fragment that is hybridizable to the complement of the promoter sequence under stringent conditions and is capable of directing expression of DNA downstream of the fragment in P. chrysogenum. Also a process for promoting expression of a coding sequence of interest in a microorganism using the isolated DNA and a process to block expression of a gene of interest in a microorganism using the isolated DNA are disclosed.
Type:
Grant
Filed:
August 2, 2000
Date of Patent:
May 6, 2003
Assignee:
Antibioticos, S.A.
Inventors:
Jose Luis Barredo Fuente, Marta Rodriguez Saiz, Alfonso J. Collados de la Vieja, Migeul Angel Moreno Valle, Francisco Salto Maldonado, Bruno Diez Garcia
Abstract: A method for selecting compounds capable of modulating protein-protein interactions is provided, in which two fusion proteins are prepared and allowed to interact with each other in the presence of test compounds. The interaction between the two fusion proteins leads to protein trans-splicing, producing an active reporter. Compounds that disrupt or enhance protein-protein interactions can be selected based on the presence or absence of the active reporter.
Abstract: The present invention relates, in general, to a novel genus of bacteria known as Ketogulonigenium. The present invention further relates to transformed Ketogulonigenium, and methods of transforming Ketogulonigenium. The present invention also relates to nucleic acid molecules, and vectors.
Abstract: The present invention relates to a process for the production of 2-keto-L-gulonic acid by fermentative conversion of L-sorbose and/or D-sorbitol. The present invention further relates to novel bacterial strains useful in this process.
Type:
Grant
Filed:
October 10, 2000
Date of Patent:
April 1, 2003
Assignee:
Archer-Daniels-Midland Company
Inventors:
Steven F. Stoddard, Hungming J. Liaw, John Eddington, Yueqin Yang
Abstract: Disclosed is a recombinant slow-growing mycobacterium comprising at least one mycobacterial gene containing an unmarked mutation, where an “unmarked mutation” is a mutated nucleotide sequence introduced into a mycobacterium where the introduced mutated nucleotide sequence does not contain a selectable marker, such as a gene conferring antibiotic resistance to the recombinant mycobacterium incorporating the mutated nucleotide sequence. Also disclosed is a method for preparing a recombinant slow-growing mycobacterium comprising at least one mycobacterial gene containing an unmarked mutation, as well as a vaccine comprising a recombinant slow-growing mycobacterium having at least one mycobacterial gene containing an unmarked mutation dispersed in a physiologically acceptable carrier. Further disclosed is a method of treating or preventing tuberculosis in a subject comprising administering the vaccine of the present invention in an amount effective to treat or prevent tuberculosis in the subject.
Abstract: The present invention relates to polynucleotide molecules comprising nucleotide sequences encoding an aveC gene product, which polynucleotide molecules can be used to alter the ratio or amount of class 2:1 avermectins produced in fermentation cultures of S. avermitilis. The present invention further relates to vectors, host cells, and mutant strains of S. avermitilis in which the aveC gene has been inactivated, or mutated so as to change the ratio or amount of class 2:1 avermectins produced.
Type:
Grant
Filed:
January 22, 2001
Date of Patent:
January 28, 2003
Assignees:
Pfizer, Inc., Pfizer Products, Inc.
Inventors:
Kim J. Stutzman-Engwall, Hamish McArthur, Yoshihiro Katoh
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 present invention provides methods of introducing a polynucleotide into a target cell, wherein the method employs a light generating protein coding sequence acting as a reporter. An important advantage of the methods described herein is that drug resistant target cells or target cells having no useful auxotrophic markers can be effectively transformed. The present invention also includes transformed cells produced by the methods described herein. Also described are light generating protein coding sequence modifications, a variety of vectors, and methods of using the transformed cells of the present invention.
Type:
Application
Filed:
March 7, 2002
Publication date:
December 19, 2002
Inventors:
Kevin P. Francis, Timothy C. Doyle, Kevin A. Nawotka
Abstract: A method of efficiently sequencing multiple exons from complex genomic DNAs is disclosed. The methodology includes the use of bacterial and bacteriophage-derived artificial chromosomes (BBPACS) in novel gene trapping protocols. Targeted gene trapping by homologous recombination, and random gene trapping with the use of a transposon system are exemplified. Included in the invention are methods of preparing a gene map from BBPAC contigs, the resulting gene maps, methods of constructing a cDNA library from BBPAC contigs, and the resulting cDNA libraries.
Type:
Grant
Filed:
August 24, 2000
Date of Patent:
November 26, 2002
Assignee:
The Rockefeller University
Inventors:
Nathaniel Heintz, Weining Jiang, Xiangdong W. Yang
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.
Type:
Grant
Filed:
April 21, 1999
Date of Patent:
November 19, 2002
Assignee:
Council of Scientic and Industrial Research
Abstract: The aim of the invention is to use at least one conditional promoter to implement a method for detecting one or more proteins interacting directly or indirectly with at least two other proteins during the formation of a protein complex, or during the inhibition of the formation of a protein complex.
Type:
Grant
Filed:
February 23, 2000
Date of Patent:
November 12, 2002
Assignees:
Institut National de la Sante et de la Recherche Medicale
(Inserm), Centre National de la Recherche Scientifique, Bristol-Myers Squibb Company
Inventors:
Jacques Camonis, Francisco Romero-Portillo, Ricardo Attar, Jean-Marc Egly, Franck Tirode
Abstract: The present invention provides a method for generating and isolating cell lines that functionally express molecular targets for drug discovery without utilizing information from the nucleic acid or amino acid sequence of the target protein. This procedure for the first time allows one to develop fast, high throughput screens for evaluation of test compounds that may modulate molecular targets whose specific nucleic acid or amino acid sequences are unavailable.
Type:
Grant
Filed:
August 10, 1999
Date of Patent:
October 29, 2002
Assignee:
Bristol-Myers Squibb Company
Inventors:
C. David Weaver, Todd A. Verdoorn, Gordon W. Robinson
Abstract: A method for expressing and purifying foreign proteins in photosynthetic organisms comprising the simultaneous expression of both the heterologous protein and a means for compartmentalizing or sequestering of the protein.
Abstract: Organisms are provided which express enzymes such as glycerol dehydratase, diol dehydratase, acyl-CoA transferase, acyl-CoA synthetase &bgr;-ketothiolase, acetoacetyl-CoA reductase, PHA synthase, glycerol-3-phosphate dehydrogenase and glycerol-3-phosphatase, which are useful for the production of PHAs. In some cases one or more of these genes are native to the host organism and the remainder are provided from transgenes. These organisms produce poly(3-hydroxyalkanoate) homopolymers or co-polymers incorporating 3-hydroxypropionate or 3-hydroxyvalerate monomers wherein the 3-hydroxypropionate and 3-hydroxyvalreate units are derived from the enzyme catalysed conversion of diols. Suitable diols that can be used include 1,2-propanediol, 1,3 propanediol and glycerol. Biochemical pathways for obtaining the glycerol from normal cellular metabolites are also described.
Abstract: The invention relates to improved E. coli bacteria with enhanced viability at low temperatures, methods for producing improved bacterial strains capable of enhanced viability at low temperatures, and the isolation and use of genetic material capable of enhancing the viability of bacteria at low temperatures. In addition to the enhanced viability at low temperatures, the bacteria may exhibit enhanced transformation efficiencies after storage at low temperatures. As such, the invention may be used for the insertion of exogenous DNA sequences into the bacteria of the invention.
Type:
Application
Filed:
January 8, 1999
Publication date:
September 26, 2002
Inventors:
FREDRIC R. BLOOM, JONATHAN KUO, JHY-JHU LIN, JIN MA
Abstract: Compositions and methods for targeted genetic manipulation of an organism are provided. The compositions are novel integration vectors derived from the Mu bacteriophage comprising an active cleaved donor complex (CDC) and further comprising a targeting mechanism whereby integration of the Mu transposable cassette may be directed to a predetermined target site within a host organism's genome. These integration vectors comprise a Mu transposable cassette and one or more navigator elements that direct targeted insertion of the CDC. Methods of the invention utilize the integration vectors of the invention to insert the Mu transposable cassette into a target site of an organism's genome. This insertion occurs in the absence of the MuB accessory protein. The methods are useful for modulating activity of known genes and for targeting integration of nucleotide sequences of interest into a specific location of an organism's genome. Accordingly, the methods may also be used to create gene disruptions and knockouts.
Abstract: The invention provides a method for making a cell which does not contain a natural nisA gene but expresses a variant nisA gene, said method comprising the steps of providing a cell that contains a natural, chromosomal nisA gene and substituting the natural nisA gene or part thereof with a variant nisA gene at the chromosomal location of the natural nisA gene or part thereof. The invention further provided a process for producing variant nisin and cells for use in the same.
Abstract: The present invention relates to isolated Porphorymonas gingivalis polynucleotides. The polynucleotides comprises a contiguous sequence of at least 20 nucleotides which is identical to a contiguous sequence of at least 20 nucleotides within a sequence selected from the group consisting of SEQ. ID. NO. 1 to SEQ. ID. NO. 1120 and sequences complementary thereto.
Abstract: The invention involves viral vectors that can be used to transduce a target cell, i.e., to introduce genetic material into the cell. The targets of interest are eukaryotic cells and particularly human cells. The transduction can be done in vivo or in vitro. More particularly the invention concerns viral vectors that have chimeric envelope proteins and contain the IgG-binding domain of protein A. These vectors when used in conjunction with antibodies targeting a particular cell are particularly useful for gene therapy.
Abstract: Disclosed are methods for inhibiting angiogenesis using cyclin dependent kinase inhibitors (CDKi) and fusion proteins thereof, recombinant viruses comprising transgenes and nucleic acid sequences encoding the same, and liposomes carrying the same as angiogenesis-inhibiting reagents.
Type:
Grant
Filed:
December 9, 1999
Date of Patent:
July 16, 2002
Assignees:
Cell Genesys, Inc., GPC Biotech Inc.
Inventors:
Salil Patel, James McArthur, Jeno Gyuris
Abstract: Novel cell lines useful for trans-complementing E1-deleted adenoviral vectors are described. The cell lines are capable of providing high yields of E1-deleted adenoviral vectors in the absence of replication-competent adenovirus over multiple passages.
Type:
Application
Filed:
January 16, 2002
Publication date:
July 11, 2002
Applicant:
The Trustees of the University of Pennsylvania
Abstract: The present invention relates to a process for the production of polyhydroxyoctanoate, said method involving construction of a multifunctional Escherichia coli-Streptomyces conjugative shuttle vector, development of a recombinant vector designated as pCAB218, which is used to transform Streptomyces lividans TK64, such that it is capable of producing polyhydroxyoctanoate (PHO) in substantial amounts when grown in a conventional mineral medium.
Abstract: The present invention provides compositions and methods of producing recombinant AAV (rAAV) virions in large amounts or high titers. Also provided are methods for producing stably transformed host cells capable of producing rAAV virions.
Abstract: A method is provided for detecting a presence of HIV virus in a sample comprising: taking a culture of recombinant cells which (a) are capable of cell division, (b) express CD4 receptor and one or more additional cell surface receptors necessary to allow the HIV virus to infect, (c) enable the HIV virus to replicate and infect the noninfected cells in the cell culture, and (d) comprise a reporter sequence introduced into the recombinant cells comprising a reporter gene whose expression is regulated by a protein specific to HIV viruses which is expressed from a genome of an HIV virus upon infection of the recombinant cell by the HIV virus; contacting the cell culture with a sample to be analyzed for the presence of HIV virus in the sample; and detecting a change in a level of expression of the reporter gene in cells in the recombinant cell culture.
Type:
Grant
Filed:
May 18, 1999
Date of Patent:
June 18, 2002
Assignee:
MUSC Foundation for Research Development
Abstract: DNA fragments which contain a sequence of DNA which encodes a protective peptide-fused &agr;-hANP in which the protective peptide has a C-terminus lysine residue which is directly fused to the N-terminus of the &agr;-hANP, vectors which contain such a DNA sequence, and microorganisms transformed which such a vector are useful for the production of &agr;-hANP.
Abstract: The sequences of nucleic acids encoding proteins required for E. coli proliferation are disclosed. The nucleic acids can be used to express proteins or portions thereof, to obtain antibodies capable of specifically binding to the expressed proteins, and to use those expressed proteins as a screen to isolate candidate molecules for rational drug discovery programs. The nucleic acids can also be used to screen for homologous genes that are required for proliferation in microorganisms other than E. coli. The nucleic acids can also be used to design expression vectors and secretion vectors. The nucleic acids of the present invention can also be used in various assay systems to screen for proliferation required genes in other organisms as well as to screen for antimicrobial agents.
Type:
Application
Filed:
July 23, 2001
Publication date:
April 18, 2002
Applicant:
ELITRA PHARMACEUTICALS, INC.
Inventors:
Judith Zyskind, Kari L. Ohlsen, John Trawick, R. Allyn Forsyth, Jamie M. Froelich, Grant J. Carr, Robert T. Yamamoto, H. Howard Xu
Abstract: Novel cell lines useful for trans-complementing E1-deleted adenoviral vectors are described. The cell lines are capable of providing high yields of E1-deleted adenoviral vectors in the absence of replication-competent adenovirus over multiple passages.
Type:
Grant
Filed:
September 11, 2000
Date of Patent:
April 2, 2002
Assignee:
The Trustees of the University of Pennsylvania
Abstract: The present invention relates, in general, to a novel genus of bacteria known as Ketogulonigenium. The present invention further relates to transformed Ketogulonigenium, and methods of transforming Ketogulonigenium. The present invention also relates to nucleic acid molecules, and vectors.
Abstract: This invention relates to an isolated nucleic acid fragment encoding a sulfate assimilation protein. The invention also relates to the construction of a chimeric gene encoding all or a portion of the sulfate assimilation protein, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the sulfate assimilation protein in a transformed host cell.
Type:
Grant
Filed:
July 1, 1999
Date of Patent:
January 15, 2002
Assignee:
E. I. du Pont de Nemours & Company
Inventors:
Saverio Carl Falco, Stephen M. Allen, Shawn L. Anderson, J. Antoni Rafalski
Abstract: Methods for the controllable expression in high yields of heterologous proteins under the control of a modified tryptophan promoter-operator system are disclosed. The system employed involves sequential resort to tryptophan repression and derepression, as well as deletion of an attenuator function native to the tryptophan operon. Means are also disclosed for effecting deletions at any given point within a gene fragment and for the ready identification of transformant bacteria via triple ligation procedures.
Type:
Grant
Filed:
June 6, 1995
Date of Patent:
December 25, 2001
Assignee:
Genentech, Inc.
Inventors:
Dennis G. Kleid, Daniel G. Yansura, Herbert L. Heyneker, Giuseppe F. Miozzari
Abstract: The present invention provides for an isolated nucleic acid molecule encoding a light chain protein of an antibody, wherein the antibody binds specifically to a protein specifically recognized by monoclonal antibody 5c8 produced by the hybridoma having ATCC Accession Number HB 10916. The invention also provides for an isolated nucleic acid molecule encoding a heavy chain protein of an antibody, wherein the antibody binds specifically to a protein specifically recognized by monoclonal antibody 5c8 produced by the hybridoma having ATCC Accession Number HB 10916. The present invention also provides for a gene transfer vector comprising a nucleic acid molecule, a host vector system comprising the gene transfer vector, and a composition comprising a nucleic acid molecule.
Type:
Grant
Filed:
April 7, 1999
Date of Patent:
December 18, 2001
Assignee:
The Trustees of Columbia University in the City of New
York
Inventors:
Seth Lederman, Leonard Chess, Michael J. Yellin
Abstract: DNA encoding a therapeutically suitable glutaminase has been molecularly cloned. This allows one to obtain a polypeptide which is a therapeutically suitable glutaminase free of contaminating endotoxin. It has been found that this polypeptide is a potent anti-viral agent and when coupled to an anti-tumor monoclonal antibody is a potent anti-cancer agent. The glutaminase of the present invention is particularly useful for treating lung, breast and colon cancer cells and in the treatment of HIV-infected cells.
Type:
Grant
Filed:
April 25, 1995
Date of Patent:
November 6, 2001
Assignee:
ME Medical Enzymes AG
Inventors:
Joseph Roberts, Thomas W MacAllister, Natarajan Sethuraman, Abbie G. Freeman
Abstract: The present invention relates to enzymes produced by mutating the genes for a number of subtilases and expressing the mutated genes in suitable hosts are presented. The enzymes exhibit improved autoproteolytic stability in comparison to their wild type parent enzymes.
Type:
Grant
Filed:
November 4, 1997
Date of Patent:
October 9, 2001
Assignee:
Novozymes A/S
Inventors:
Claus von der Osten, Torben Halkier, Carsten Andersen, Peter Bauditz, Peter Kamp Hansen
Abstract: Tools for genetically engineering Pasteurellaceae are provided. Replication-conditional plasmids which are useful for the Pasteurellaceae have been isolated and characterized. The plasmids can be utilized for delivery of DNA segments into the Pasteurellaceae in situations where control of extra-chromosomal replication desired, such as in achieving allelic exchange or site-directed mutagenesis.
A restriction endonuclease, HsoI, was isolated from a bovine lung isolate of Haemophilus somnus. The enzyme was found to be a true isoschizomer of HinPI, a commercially available enzyme originally isolated from Haemophilus influenzae PI. Commercially available HhaI methyl transferase was found to protect against cleavage by both enzymes. Methylation of foreign plasmid DNA was found to enhance transformation of Haemophilus somnus in excess of four orders of magnitude.
Type:
Grant
Filed:
November 21, 2000
Date of Patent:
March 11, 2003
Assignees:
Biotechnology Research and Development Corporation, The United States of America as represented by the Department
of Agriculture