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WO1999061651A3 - Methods for producing a polypeptide by modifying the copy number of a gene - Google Patents

Methods for producing a polypeptide by modifying the copy number of a gene Download PDF

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Publication number
WO1999061651A3
WO1999061651A3 PCT/US1999/011816 US9911816W WO9961651A3 WO 1999061651 A3 WO1999061651 A3 WO 1999061651A3 US 9911816 W US9911816 W US 9911816W WO 9961651 A3 WO9961651 A3 WO 9961651A3
Authority
WO
WIPO (PCT)
Prior art keywords
nucleic acid
polypeptide
acid sequence
cell
methods
Prior art date
Application number
PCT/US1999/011816
Other languages
French (fr)
Other versions
WO1999061651A2 (en
Inventor
Debbie S Yaver
Original Assignee
Novo Nordisk Biotech Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Novo Nordisk Biotech Inc filed Critical Novo Nordisk Biotech Inc
Priority to JP2000551031A priority Critical patent/JP2002516116A/en
Priority to EP99925956A priority patent/EP1080210A2/en
Priority to AU42139/99A priority patent/AU4213999A/en
Publication of WO1999061651A2 publication Critical patent/WO1999061651A2/en
Publication of WO1999061651A3 publication Critical patent/WO1999061651A3/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/67—General methods for enhancing the expression
    • C12N15/69—Increasing the copy number of the vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90—Stable introduction of foreign DNA into chromosome
    • C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
    • C12N9/20—Triglyceride splitting, e.g. by means of lipase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01004—Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Mycology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a mutant cell under conditions conducive for production of the polypeptide, wherein (i) the mutant cell is related to a parent cell, which parent cell comprises at least two tandem copies of a nucleic acid sequence encoding the polypeptide or a nucleic acid sequence encoding the polypeptide, which nucleic acid sequence comprises repeat sequences at the 5' and 3' ends of the nucleic acid sequence, by the introduction of a nucleic acid construct into the genome of the parent cell at a locus which is within or not within the nucleic acid sequence(s), wherein the introduction of the nucleic acid construct into the locus modifies the copy number of the nucleic acid sequence(s) and the modification of the copy number is not a result of selective pressure; and (ii) the mutant cell produces more or less of the polypeptide than the parent cell when both cells are cultivated under the same conditions; and (b) recovering the poylpeptide from the cultivation medium. The present invention also relates to methods for obtaining a mutant cell and mutant cells.
PCT/US1999/011816 1998-05-27 1999-05-27 Methods for producing a polypeptide by modifying the copy number of a gene WO1999061651A2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
JP2000551031A JP2002516116A (en) 1998-05-27 1999-05-27 Method for producing a polypeptide by changing the copy number of a gene
EP99925956A EP1080210A2 (en) 1998-05-27 1999-05-27 Methods for producing a polypeptide by modifying the copy number of a gene
AU42139/99A AU4213999A (en) 1998-05-27 1999-05-27 Methods for producing a polypeptide by modifying the copy number of a gene

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US8550598A 1998-05-27 1998-05-27
US09/085,505 1998-05-27

Publications (2)

Publication Number Publication Date
WO1999061651A2 WO1999061651A2 (en) 1999-12-02
WO1999061651A3 true WO1999061651A3 (en) 2000-03-02

Family

ID=22192064

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1999/011816 WO1999061651A2 (en) 1998-05-27 1999-05-27 Methods for producing a polypeptide by modifying the copy number of a gene

Country Status (5)

Country Link
EP (1) EP1080210A2 (en)
JP (1) JP2002516116A (en)
CN (1) CN1307642A (en)
AU (1) AU4213999A (en)
WO (1) WO1999061651A2 (en)

Families Citing this family (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2755300A (en) * 1999-02-02 2000-08-25 Novozymes Biotech, Inc. Methods for producing polypeptides in fungal cells
CN1312260A (en) * 2000-03-07 2001-09-12 上海博德基因开发有限公司 Human proteinase regulating protein 10 as one new polypeptide and polynucleotides encoding this polypeptide
AU2002225856A1 (en) * 2000-11-06 2002-05-15 Pepgen Corporation Method of determining gene copy number
EP1499733B1 (en) 2002-04-22 2010-03-10 Novozymes, Inc. Methods for preparing variants of a dna sequence in filamentous fungi
EP2228440B1 (en) 2003-05-02 2012-08-22 Novozymes Inc. Variants of beta-glucosidases
RU2340665C2 (en) * 2003-08-22 2008-12-10 Сантори Лимитед Method of selecting lipid-producing mushrooms
ES2538824T3 (en) 2003-08-25 2015-06-24 Novozymes, Inc. Variants of glycoside hydrolases
CN100543141C (en) * 2003-09-16 2009-09-23 周琪 Method for producing gene engineering recombination 192 peptide human growth hormone
ES2437198T3 (en) 2003-10-28 2014-01-09 Novozymes Inc. Polypeptides with beta-glucosidase activity and isolated polynucleotides encoding the polypeptides
WO2005067531A2 (en) 2004-01-16 2005-07-28 Novozymes Inc. Methods for degrading lignocellulosic materials
CN104726431B (en) 2004-01-30 2018-06-01 诺维信股份有限公司 Polypeptide and its coded polynucleotide with cellulolytic enhancing activity
CA2565888C (en) 2004-05-27 2014-10-14 Novozymes, Inc. Methods for transforming and expression screening of filamentous fungal cells with a dna library
US7824884B2 (en) 2005-04-27 2010-11-02 Novozymes, Inc. Polypeptides having endoglucanase activity and polynucleotides encoding same
NZ589570A (en) 2005-09-30 2012-06-29 Novozymes Inc Methods for enhancing the degradation or conversion of cellulosic material
WO2007115201A2 (en) 2006-03-30 2007-10-11 Novozymes, Inc. Polypeptides having endoglucanase activity and polynucleotides encoding same
FR2927089B1 (en) * 2008-02-05 2011-03-25 Inst Nat De La Rech Agronomique Inra METHOD OF TARGETED INTEGRATION OF MULTICOPIES OF A GENE OF INTEREST IN A YARROWIA STRAIN
EP2358878B1 (en) 2008-11-20 2014-10-15 Novozymes Inc. Polypeptides having amylolytic enhancing activity and polynucleotides encoding same
US8338121B2 (en) 2008-12-19 2012-12-25 Novozymes, Inc. Methods for determining cellulolytic enhancing activity of a polypeptide
CN102388134A (en) 2009-01-28 2012-03-21 诺维信股份有限公司 Polypeptides having beta-glucosidase activity and polynucleotides encoding same
US8563268B2 (en) 2009-03-17 2013-10-22 Novozymes A/S Polypeptide having tyrosinase activity
WO2011050037A1 (en) 2009-10-23 2011-04-28 Novozymes, Inc. Cellobiohydrolase variants and polynucleotides encoding same
WO2011057140A1 (en) 2009-11-06 2011-05-12 Novozymes, Inc. Compositions for saccharification of cellulosic material
DK2496693T3 (en) 2009-11-06 2018-01-22 Novozymes Inc Polypeptides with cellobiohydrolase activity and polynucleotides encoding them
WO2011057083A1 (en) 2009-11-06 2011-05-12 Novozymes, Inc. Polypeptides having xylanase activity and polynucleotides encoding same
US10246691B2 (en) 2010-09-30 2019-04-02 Novozymes, Inc. Variants of polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
DK2622070T3 (en) 2010-09-30 2016-11-21 Novozymes Inc Variants of polypeptides having cellulolytic enhancing ACTIVITY AND POLYNUCLEOTIDES ENCODING THEM
EP3023492B1 (en) 2010-10-01 2018-01-24 Novozymes, Inc. Beta-glucosidase variants and polynucleotides encoding same
CN103339252A (en) 2010-11-18 2013-10-02 诺维信股份有限公司 Chimeric polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
WO2012103350A1 (en) 2011-01-26 2012-08-02 Novozymes A/S Polypeptides having cellobiohydrolase activity and polynucleotides encoding same
EP2691519A1 (en) 2011-03-31 2014-02-05 Novozymes, Inc. Cellulose binding domain variants and polynucleotides encoding same
US9926547B2 (en) 2011-04-28 2018-03-27 Novozymes, Inc. Polypeptides having endoglucanase activity and polynucleotides encoding same
US9624518B2 (en) 2011-04-29 2017-04-18 Novozymes, Inc. Methods for enhancing the degradation or conversion of cellulosic material
BR112014007651A2 (en) 2011-09-30 2017-04-11 Novozymes Inc isolated chimeric polypeptide, isolated polynucleotide, methods for producing a chimeric polypeptide and a fermentation product, for degrading or converting a cellulosic material, and for fermenting a transgenic cellulosic material, plant, plant part or cell, and broth formulation whole or cell culture composition
BR112014015228B1 (en) 2011-12-20 2022-07-05 Novozymes, Inc. GENITOR CELOBIOHIDROLASE VARIANT, PROCESSES FOR DEGRADING OR CONVERTING A CELLULOSIC MATERIAL, FOR PRODUCTION OF A FERMENTATION PRODUCT, AND FOR FERMENTATION OF A CELLULOSIC MATERIAL, WHOLE BROTH FORMULATION OR CELL CULTURE COMPOSITION, AND, TRANSGENIC MICROBIAN HOST CELL
CN117716046A (en) * 2021-06-29 2024-03-15 诺维信公司 Process for producing fermentation products using fiber-degrading enzymes in fermentation

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2563533A1 (en) * 1984-04-27 1985-10-31 Centre Nat Rech Scient METHOD FOR AMPLIFYING EXPRESSION OF A GENE DETERMINED IN BACILLUS SUBTILIS AND OBTAINED STRAINS
WO1988003169A1 (en) * 1986-10-27 1988-05-05 Whitehead Institute For Biomedical Research Gene amplification using retrotransposons
EP0343783A2 (en) * 1988-04-22 1989-11-29 Btg Usa Inc Method for producing cells containing stably integrated foreign DNA at a high copy number, the cells produced by this method, and the use of these cells to produce the polypeptides coded for by the foreign DNA
WO1991009955A1 (en) * 1989-12-22 1991-07-11 Applied Research Systems Ars Holding N.V. Endogenous gene expression modification with regulatory element
WO1992001800A1 (en) * 1990-07-20 1992-02-06 Chiron Corporation Method for integrative transformation of yeast using dispersed repetitive elements
WO1995031560A1 (en) * 1994-05-13 1995-11-23 Transkaryotic Therapies, Inc. Dna construct for effecting homologous recombination and uses thereof
WO1998011203A1 (en) * 1996-09-13 1998-03-19 Novo Nordisk Biotech, Inc. Cells having dna insertion mutations which produce altered amounts of a polypeptide

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2563533A1 (en) * 1984-04-27 1985-10-31 Centre Nat Rech Scient METHOD FOR AMPLIFYING EXPRESSION OF A GENE DETERMINED IN BACILLUS SUBTILIS AND OBTAINED STRAINS
WO1988003169A1 (en) * 1986-10-27 1988-05-05 Whitehead Institute For Biomedical Research Gene amplification using retrotransposons
EP0343783A2 (en) * 1988-04-22 1989-11-29 Btg Usa Inc Method for producing cells containing stably integrated foreign DNA at a high copy number, the cells produced by this method, and the use of these cells to produce the polypeptides coded for by the foreign DNA
WO1991009955A1 (en) * 1989-12-22 1991-07-11 Applied Research Systems Ars Holding N.V. Endogenous gene expression modification with regulatory element
WO1992001800A1 (en) * 1990-07-20 1992-02-06 Chiron Corporation Method for integrative transformation of yeast using dispersed repetitive elements
WO1995031560A1 (en) * 1994-05-13 1995-11-23 Transkaryotic Therapies, Inc. Dna construct for effecting homologous recombination and uses thereof
WO1998011203A1 (en) * 1996-09-13 1998-03-19 Novo Nordisk Biotech, Inc. Cells having dna insertion mutations which produce altered amounts of a polypeptide

Also Published As

Publication number Publication date
AU4213999A (en) 1999-12-13
WO1999061651A2 (en) 1999-12-02
EP1080210A2 (en) 2001-03-07
JP2002516116A (en) 2002-06-04
CN1307642A (en) 2001-08-08

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