DK2683732T3 - Vektor-vært-system - Google Patents

Vektor-vært-system Download PDF

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Publication number
DK2683732T3
DK2683732T3 DK12719604.6T DK12719604T DK2683732T3 DK 2683732 T3 DK2683732 T3 DK 2683732T3 DK 12719604 T DK12719604 T DK 12719604T DK 2683732 T3 DK2683732 T3 DK 2683732T3
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Denmark
Prior art keywords
vector
host cell
gene
interest
jak
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DK12719604.6T
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English (en)
Inventor
Roelof Ary Lans Bovenberg
Jan Andries Kornelis Willem Kiel
Thibaut José Wenzel
Alrik Pieter Los
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Dsm Ip Assets Bv
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
    • C07K14/385—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from Penicillium
    • 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
    • C12N15/65—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression using markers

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

Claims (14)

1. Fremgangsmåde til fremstilling af en biologisk forbindelse af interesse, der omfatter dyrkning af en trådformig svampeværtscelle, der er deficient i et essentielt gen og omfatter en vektor, hvilken vektor omfatter mindst det essentielle gen og en autonom replikationssekvens, hvor værtscellen omfatter en rekombinant polynukleotidkonstruktion, der omfatter et polynukleotid, der koder for en biologisk forbindelse af interesse eller en forbindelse, der er inddraget i syntesen af en biologisk forbindelse af interesse, under betingelser, der bidrager til fremstillingen af den biologiske forbindelse af interesse, og isolering af forbindelsen af interesse fra dyrkningsbouillonen, hvor en rekombinant polynukleotidkonstruktion, der koder for den biologiske forbindelse af interesse eller en forbindelse, der er inddraget i syntesen af den biologiske forbindelse af interesse, er placeret på vektoren.
2. Fremgangsmåde til fremstilling af en biologisk forbindelse af interesse, der omfatter frembringelse af en trådformig svampeværtscelle ifølge en fremgangsmåde, der omfatter: a. tilvejebringelse af en trådformig svampeværtscelle og en vektor, der omfatter mindst et gen, der er essentielt for værtscellen, og en autonom replikationssekvens, hvor en rekombinant polynukleotidkonstruktion, der koder for forbindelsen af interesse eller en forbindelse, der er inddraget i syntesen af den biologiske forbindelse af interesse, er placeret på vektoren; og b. co-transformering af værtscellen med vektoren og en ødelæggelseskonstruktion for det essentielle gen til at gøre værtscellen deficient i det essentielle gen; dyrkning af en sådan værtscelle under betingelser, der bidrager til fremstillingen af den biologiske forbindelse af interesse, og isolering af forbindelsen af interesse fra dyrkningsbouillonen.
3. Fremgangsmåde ifølge krav 1 eller 2, hvor værtscellen fortrinsvis er inducerbart forøget i dens effektivitet af homolog rekombination (HR), reduceret i dens effektivitet af ikke-homolog rekombination (NHR) eller reduceret i dens forhold mellem ikke-homolog rekombination/homolog rekombination (NHR/HR).
4. Fremgangsmåde ifølge et hvilket som helst af ovennævnte krav, hvor værtscelledeficiensen i det essentielle gen er inducerbar.
5. Fremgangsmåde ifølge et hvilket som helst af ovennævnte krav, hvor det essentielle gen er et essentielt gen i svampe.
6. Fremgangsmåde ifølge et hvilket som helst af ovennævnte krav, hvor det essentielle gen er tif35- eller aurl-genet.
7. Fremgangsmåde ifølge et hvilket som helst af ovennævnte krav, hvor værtscellen er en Aspergillus, Chrysosporium, Penicillium, Rasamsonia, Talaromyces eller Trichoderma.
8. Fremgangsmåde ifølge krav 7, hvor værtscellen er Aspergillus niger, Penicillium chrysogenum eller Rasamsonia emersonii.
9. Fremgangsmåde ifølge et hvilket som helst af ovennævnte krav, hvor vektoren indeholder kontrolsekvenser fra en anden art end værtsarten.
10. Fremgangsmåde ifølge et hvilket som helst af ovennævnte krav, hvor den biologiske forbindelse af interesse er en biopolymer eller en metabolit.
11. Fremgangsmåde ifølge krav 10, hvor biopolymeren er et polypeptid.
12. Fremgangsmåde ifølge krav 11, hvor polypeptidet er et antistof eller dele deraf, et antigen, en koagulationsfaktor, et enzym, et hormon eller en hormonvariant, en receptor eller dele deraf, et regulatorprotein, et strukturprotein, en reporter eller et transportprotein, et protein, der er inddraget i en sekretionsproces, et protein, der er inddraget i en foldningsproces, en chaperone, en peptidaminosyretransporter, en glycosyleringsfaktor, en transkriptionsfaktor, et syntetisk peptid eller oligopeptid, et intracellulært protein.
13. Fremgangsmåde ifølge krav 12, hvor enzymet er en protease, ceramidase, epoxidhydrolase, aminopeptidase, acylase, aldolase, hydroxylase, aminopeptidase, lipase, non-ribosomal synthetase, polyketidsynthetase, oxidoreductase, transferase, hydrolase, lyase, isomerase, catalase, cellulase, chitinase, cutinase, deoxyribonuclease, dextranase, esterase, carbohydrase, f.eks. cellulase, såsom endoglucanase, β-glucanase, cellobiohydrolase eller β-glucosidase, et GH61-enzym, en hemicellulase, et pektinolytisk enzym, såsom xylanase, xylosidase, mannanase, galactanase, galactosidase, pektinmethylesterase, pektinlyase, pektatlyase, endopolygalacturonase, exopolygalacturonase, rhamnogalacturonase, arabanase, arabinofuranosidase, arabinoxylanhydrolase, galacturonase, lyase eller et amylolytisk enzym; hydrolase, isomerase eller ligase, phosphatase, såsom phytase, esterase, såsom lipase, phospholipase, galactolipase, et proteolytisk enzym, et mejerienzym, såsom chymosin, oxidoreductase, såsom oxidase, transferase, isomerase, phytase, aminopeptidase, asparaginase, amylase, carbohydrase, carboxypeptidase, endoprotease, metalloprotease, serinprotease, catalase, chitinase, cutinase, cyclodextrin glycosyltransferase, deoxyribonuclease, esterase, alfa-galactosidase, beta-galactosidase, glucoamylase, alfa-glucosidase, beta-glucosidase, haloperoxidase, proteindeaminase, invertase, laccase, lipase, mannosidase, mutanase, oxidase, et pektinolytisk enzym, peroxidase, phospholipase, polyphenoloxidase, ribonuclease, transglutaminase eller glucoseoxidase, hexoseoxidase, monooxygenase .
14. Fremgangsmåde ifølge et hvilket som helst af ovennævnte krav, hvor værtscellen allerede er i stand til at danne den biologiske forbindelse af interesse.
DK12719604.6T 2011-03-11 2012-03-12 Vektor-vært-system DK2683732T3 (da)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP11157871 2011-03-11
PCT/EP2012/054302 WO2012123429A1 (en) 2011-03-11 2012-03-12 Vector-host system

Publications (1)

Publication Number Publication Date
DK2683732T3 true DK2683732T3 (da) 2016-12-12

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US (1) US20140106398A1 (da)
EP (1) EP2683732B1 (da)
CN (1) CN103596970A (da)
DK (1) DK2683732T3 (da)
WO (1) WO2012123429A1 (da)

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CN104981542B (zh) 2012-03-12 2018-10-16 帝斯曼知识产权资产管理有限公司 重组系统
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EP2683732A1 (en) 2014-01-15
WO2012123429A1 (en) 2012-09-20
CN103596970A (zh) 2014-02-19
EP2683732B1 (en) 2016-08-24
US20140106398A1 (en) 2014-04-17

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