WO2001038510A3 - Integrationsvektoren und verfahren zur herstellung rekombinanter proteine in pilzen - Google Patents

Integrationsvektoren und verfahren zur herstellung rekombinanter proteine in pilzen Download PDF

Info

Publication number
WO2001038510A3
WO2001038510A3 PCT/EP2000/011687 EP0011687W WO0138510A3 WO 2001038510 A3 WO2001038510 A3 WO 2001038510A3 EP 0011687 W EP0011687 W EP 0011687W WO 0138510 A3 WO0138510 A3 WO 0138510A3
Authority
WO
WIPO (PCT)
Prior art keywords
host organism
integration
vectors
heterologous
proteins
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/EP2000/011687
Other languages
English (en)
French (fr)
Other versions
WO2001038510A2 (de
Inventor
Gerd Gellissen
Andre Diesel
Jens Klabunde
Manfred Suckow
Cornelis P Hollenberg
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Dynavax GmbH
Original Assignee
Rhein Biotech GmbH
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
Priority claimed from DE19963690A external-priority patent/DE19963690A1/de
Application filed by Rhein Biotech GmbH filed Critical Rhein Biotech GmbH
Priority to EP00985088A priority Critical patent/EP1242603B1/de
Priority to AU21621/01A priority patent/AU2162101A/en
Priority to AT00985088T priority patent/ATE264917T1/de
Priority to DE50006189T priority patent/DE50006189D1/de
Publication of WO2001038510A2 publication Critical patent/WO2001038510A2/de
Anticipated expiration legal-status Critical
Publication of WO2001038510A3 publication Critical patent/WO2001038510A3/de
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/62—Insulins
    • 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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • 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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • C12N15/815—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts for yeasts other than Saccharomyces
    • 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)

Landscapes

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

Abstract

Die Erfindung betrifft Integrationsvektoren zur gezielten und stabilen Integration heterologer DNA in das Genom eines Wirtsorganismus, insbesondere eines Pilzes, und ihre Verwendung. Ferner betrifft die Erfindung ein Verfahren zur Herstellung eines oder mehrerer rekombinanter Proteine unter Verwendung der erfindungsgemäßen Vektoren. Aufgabe der Erfindung ist es, Integrationsvektoren bereitzustellen, die es erlauben, Fremd-DNA in hoher Kopienzahl in das Genom eines Wirtsorganismus stabil zu integrieren und bei Bedarf unterschiedliche Gene für eine simultane Koproduktion verschiedener Proteine zu exprimieren. Eine weitere Aufgabe der Erfindung ist es, stabile Produktionsstämme und ein Verfahren zur Herstellung heterologer Proteine bereitzustellen. Diese Aufgabe wird durch einen Integrationsvektor gelöst, welcher: a) mindestens eine rDNA-Sequenz aus Hefe, b) mindestens ein nicht-defizientes Selektionsmarkergen und c) mindestens eine Expressionskassette umfaßt, wobei die Expressionskassette ein im Wirtsorganismus funktionsfähiges Promotorelement, einen für ein gewünschtes Protein kodierenden Bereich eines heterologen oder genuinen Genes und ein im Wirtsorganismus funktionsfähiges Terminatorelement umfaßt. Von der Erfindung wird ebenfalls ein Wirtsorganismus umfaßt, der mindestens einen erfindungsgemäßen Vektor enthält. Ein bevorzugter Wirtsorganismus ist die methylotrophe Hefe Hansenula polymorpha.
PCT/EP2000/011687 1999-11-23 2000-11-23 Integrationsvektoren und verfahren zur herstellung rekombinanter proteine in pilzen Ceased WO2001038510A2 (de)

Priority Applications (4)

Application Number Priority Date Filing Date Title
EP00985088A EP1242603B1 (de) 1999-11-23 2000-11-23 Integrationsvektoren und verfahren zur herstellung rekombinanter proteine in pilzen
AU21621/01A AU2162101A (en) 1999-11-23 2000-11-23 Vectors and method for producing recombinant proteins in fungi
AT00985088T ATE264917T1 (de) 1999-11-23 2000-11-23 Integrationsvektoren und verfahren zur herstellung rekombinanter proteine in pilzen
DE50006189T DE50006189D1 (de) 1999-11-23 2000-11-23 Integrationsvektoren und verfahren zur herstellung rekombinanter proteine in pilzen

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
DE19956297 1999-11-23
DE19956297.0 1999-11-23
DE19958327 1999-12-03
DE19958327.7 1999-12-03
DE19963690A DE19963690A1 (de) 1999-11-23 1999-12-29 Vektoren und Verfahren zur Herstellung rekombinanter Proteine in Pilzen
DE19963690.7 1999-12-29

Publications (2)

Publication Number Publication Date
WO2001038510A2 WO2001038510A2 (de) 2001-05-31
WO2001038510A3 true WO2001038510A3 (de) 2002-07-04

Family

ID=27219346

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2000/011687 Ceased WO2001038510A2 (de) 1999-11-23 2000-11-23 Integrationsvektoren und verfahren zur herstellung rekombinanter proteine in pilzen

Country Status (7)

Country Link
EP (1) EP1242603B1 (de)
AT (1) ATE264917T1 (de)
AU (1) AU2162101A (de)
DK (1) DK1242603T3 (de)
ES (1) ES2221865T3 (de)
PT (1) PT1242603E (de)
WO (1) WO2001038510A2 (de)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002053758A2 (de) * 2000-12-29 2002-07-11 Rhein Biotech Gesellschaft für neue Biotechnologische Prozesse und Produkte mbH Verfahren zum herstellen von heterologen proteinen in einem homothallischen pilz der familie sordariaceae
ITMI20011728A1 (it) * 2001-08-06 2003-02-06 Consorzio Per Le Ricerche E Lo Vettore per l'integrazione sito-specifica di sequenze di dna eterologhe in lieviti metilotrofi
EP1698702B1 (de) * 2005-03-02 2012-02-22 PharmedArtis GmbH Expression System für rekombinante Proteine
EP2279241B1 (de) * 2008-05-06 2020-04-08 Archer Daniels Midland Co. Entwicklung von stämmen der wärmetoleranten hefe hansenula polymorpha zur alkoholischen fermentierung von stärke und xylan durch expression von stärke- und xylanabbauenden enzymen
JP6475336B2 (ja) * 2014-08-21 2019-02-27 チュン−アン ユニヴァーシティ インダストリー−アカデミー コーポレーション ファウンデーションＣｈｕｎｇ−Ａｎｇ Ｕｎｉｖｅｒｓｉｔｙ Ｉｎｄｕｓｔｒｙ−Ａｃａｄｅｍｙ Ｃｏｏｐｅｒａｔｉｏｎ Ｆｏｕｎｄａｔｉｏｎ ｒＤＮＡ ＮＴＳ基盤の遺伝子多重挿入カセットセット、及びそれを用いたＧＲＡＳ等級の組換え酵母菌株

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999057279A1 (en) * 1998-05-07 1999-11-11 Doosan Corporation Plasmid for gene expression in pichia ciferrii and transformation method using the same

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999057279A1 (en) * 1998-05-07 1999-11-11 Doosan Corporation Plasmid for gene expression in pichia ciferrii and transformation method using the same

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
BERGKAMP R J M ET AL: "MULTIPLE-COPY INTEGRATION OF THE ALPHA-GALACTOSIDASE GENE FROM CYAMOPSIS TETRAGONOLOBA INTO THE RIBOSOMAL DNA OF KLUYVEROMYCES LACTIS", CURRENT GENETICS,US,NEW YORK, NY, vol. 21, no. 4/05, 1 April 1992 (1992-04-01), pages 365 - 370, XP002040755, ISSN: 0172-8083 *
BLANCAFORT PILAR ET AL: "PolI-driven integrative expression vectors for yeast.", JOURNAL OF BIOTECHNOLOGY, vol. 56, no. 1, 1997, pages 41 - 47, XP000923345, ISSN: 0168-1656 *
DALL LE M -T ET AL: "MULTIPLE-COPY INTEGRATION IN THE YEAST YARROWIA LIPOLYTICA", CURRENT GENETICS,US,NEW YORK, NY, vol. 26, no. 1, 1994, pages 38 - 44, XP002040754, ISSN: 0172-8083 *
FUJII T ET AL: "APPLICATION OF A RIBOSOMAL DNA INTEGRATION VECTOR IN THE CONSTRUCTION OF A BREWER'S YEAST HAVING ALPHA ACETOLACTATE DECARBOXYLASE ACTIVITY", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, vol. 56, no. 4, 1990, pages 997 - 1003, XP000994944, ISSN: 0099-2240 *
GELLISSEN GERD ET AL: "Application of yeasts in gene expression studies: A comparison of Saccharomyces cerevisiae, Hansenula polymorpha and Kluyveromyces lactis: A review.", GENE (AMSTERDAM), vol. 190, no. 1, 1997, pages 87 - 97, XP002164545, ISSN: 0378-1119 *
HOLLENBERG CORNELIS P ET AL: "Production of recombinant proteins by methylotrophic yeasts.", CURRENT OPINION IN BIOTECHNOLOGY, vol. 8, no. 5, 1997, pages 554 - 560, XP000939179, ISSN: 0958-1669 *
LOPES T S ET AL: "HIGH-COPY-NUMBER INTEGRATION INTO THE RIBOSOMAL DNA OF SACCHAROMYCES CEREVISIAE: A NEW VECTOR FOR HIGH-LEVEL EXPRESSION", GENE,NL,ELSEVIER BIOMEDICAL PRESS. AMSTERDAM, vol. 79, no. 2, 1989, pages 199 - 206, XP000034807, ISSN: 0378-1119 *
LOPES T S ET AL: "MECHANISM OF HIGH-COPY-NUMBER INTEGRATION OF PMIRY-TYPE VECTORS INTO THE RIBOSOMAL DNA OF SACCHAROMYCES-CEREVISIAE", GENE (AMSTERDAM), vol. 105, no. 1, 1991, pages 83 - 90, XP000986021, ISSN: 0378-1119 *
ROSSOLINI G M ET AL: "KLUYVEROMYCES-LACTIS RDNA AS A TARGET FOR MULTIPLE INTEGRATION BY HOMOLOGOUS RECOMBINATION", GENE (AMSTERDAM), vol. 119, no. 1, 1992, pages 75 - 81, XP000986022, ISSN: 0378-1119 *
YIP C W ET AL: "RIBOSOMAL RNA GENES OF ENDOMYCES FIBULIGER: ISOLATION, SEQUENCING AND THE USE OF THE 26S RRNA GENE IN INTEGRATIVE TRANSFORMATION OF SACCHAROMYCES CEREVISIAE FOR EFFICIENT EXPRESSION OF THE ALPHA-AMYLASE GENE OF ENDOMYCES FIBULIGER", WORLD JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY,GB,RAPID COMMUNICATIONS OF OXFORD, OXFORD, vol. 13, no. 1, 1997, pages 103 - 117, XP000677652, ISSN: 0959-3993 *

Also Published As

Publication number Publication date
AU2162101A (en) 2001-06-04
PT1242603E (pt) 2004-09-30
EP1242603A2 (de) 2002-09-25
ES2221865T3 (es) 2005-01-16
DK1242603T3 (da) 2004-06-28
WO2001038510A2 (de) 2001-05-31
ATE264917T1 (de) 2004-05-15
EP1242603B1 (de) 2004-04-21

Similar Documents

Publication Publication Date Title
IL152272A0 (en) Novel expression-regulating sequences and expression products in the field of filamentous fungi
CA2128589A1 (en) Selection marker gene free recombinant strains: a method for obtaining them and the use of these strains
EP1033405A3 (de) DNS-fragmente mit bestimmter Sequenz und die dadurch kodierte Polypeptide
EP2325319A3 (de) Produkt und Verfahren zur Umwandlung Thraustochytriales Mikroorganismen
Radzio et al. Synthesis of biotechnologically relevant heterologous proteins in filamentous fungi
AU2162101A (en) Vectors and method for producing recombinant proteins in fungi
EP0225078A3 (de) Zusammengesetzte Expressionskassetten für die Transformation von Pilzen
WO2000004167A3 (en) Genes encoding sulfate assimilation proteins
AU3868197A (en) Yeast vectors and process for producing proteins with the use of the same
ATE339506T1 (de) Rifamycin biosynthese genkluster
ID22037A (id) PROSES REKOMBINAN UNTUK MEMBUAT ANTIGEN MALARIA KOMPLET gp190/MSPI
DE69626494D1 (de) Promotor/terminator des formatdehydrogenase-gens von candida boidinii
WO1999051756A3 (en) Promoter and constructions for expression of recombinant proteins in filamentous fungi
WO2002081650A3 (en) Alcohol oxidase 1 regulatory nucleotide sequences for heterologous gene expression in yeast
WO2002081702A1 (fr) Genes de desaturase en position $g(d)6 d'acide gras, plasmides et transformants contenant ces genes
WO2003031596A3 (en) Production of a highly active, soluble form of the cytochrome p450 reductase (cpr a) from candida tropicalis
WO2003014363A8 (en) Vector for site-specific integration of heterologous dna sequences into the rdn a sites of methylotrophic yeasts
CA2091236A1 (en) Ornithine carbamoyl transferase gene and utilization of the dna
WO2001079489A3 (en) 39228, a human alcohol dehydrogenase and uses therefor
RU2001124653A (ru) Новая трансформационная система для экспрессии генов грибов под контролем регуляторных систем мицелиального гриба Penicillium canescnes
NO331102B1 (no) Promotor, ekspresjonsvektor og vert, og fremgangsmate for a produsere et aktuelt protein
WO2003020912A3 (en) Novel clark protein and nucleic acid molecules and uses therefor
WO2002053758A3 (de) Verfahren zum herstellen von heterologen proteinen in einem homothallischen pilz der familie sordariaceae
EP1338653A3 (de) Verbesserter Expressionsvektor für die Herstellung rekombinanter Proteine
AU2001263920A1 (en) Method for the production of proteins in a yeast of the genusarxula and suitable promoters

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 2000985088

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 1020027006604

Country of ref document: KR

AK Designated states

Kind code of ref document: A3

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A3

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

WWP Wipo information: published in national office

Ref document number: 2000985088

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWP Wipo information: published in national office

Ref document number: 1020027006604

Country of ref document: KR

WWG Wipo information: grant in national office

Ref document number: 2000985088

Country of ref document: EP

WWG Wipo information: grant in national office

Ref document number: 1020027006604

Country of ref document: KR