FI114481B - Selektionsmarkörgenfria trådformiga svampstammar, förfarande för deras framställning, och användning av dessa stammar - Google Patents
Selektionsmarkörgenfria trådformiga svampstammar, förfarande för deras framställning, och användning av dessa stammar Download PDFInfo
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- FI114481B FI114481B FI943485A FI943485A FI114481B FI 114481 B FI114481 B FI 114481B FI 943485 A FI943485 A FI 943485A FI 943485 A FI943485 A FI 943485A FI 114481 B FI114481 B FI 114481B
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- 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
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
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- 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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- Genetics & Genomics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
Claims (25)
1. Vektor, kännetecknad därav, att den innehäller en dominerande, dubbelriktad och kontraselektiv 5 selektionsmarkörgen, som är flankerad av DNA- upprepningssekvenser, som möjliggör inre rekombination i genomet av en trädformig svamp.
2. Vektor enligt patentkrav 1, kännetecknad därav, att DNA-10 upprepningssekvenserna väljs frän 5'- eller 3'-ändan av DNA- sekvensen som ska avslägnas.
3. Vektor enligt patentkravet 1 eller 2, kännetecknad därav, att selektionsmarkörgenen är en acetamidasgen. 15
4. Vektor enligt patentkrav 3, kännetecknad därav, att acetamidasgenen härstammar frän en svamp.
5. Vektor enligt patentkrav 4, kännetecknad därav, att 20 acetamidasgenen är frän arten Aspergillus.
6. Vektor enligt nägot av patentkraven 1-5, kännetecknad därav, att vektorn innehäller ocksä ett önskat DNA-fragment. f » » « l ·
7. Vektor enligt patentkrav 6, kännetecknad därav, att det » ··· önskade DNA-f ragment et innehäller nägot av följande genetiska ·. element eller deras kombinationer: en gen, en cDNA, en promotor, en terminator, ett regleringselement, en intron, en igenkänningssekvens för ett DNA-bindande protein, ett 30 translationsinitieringsställe eller ett restriktionsställe.
' · · : 8. Vektor enligt patentkrav 7, kännetecknad därav, att det önskade DNA-fragmentet innehäller en sekvens som kodar för kymosin, fytas, xylanas, lipas, amylas, proteas eller β-35 galaktosidas.
9. Rekombinant trädformig svamp som är transformerad med en vektor enligt nägot av patentkraven 1-8. / 114481
10. Trädformig svamp enligt patentkrav 9, kännetecknad därav, att den omfattar minst tva olika vektorer enligt nagot av patentkraven 1-8. 5
11. Trädformig svamp enligt patentkrav 9 eller 10, kännetecknad därav, att vektorn har integrerats i genomet av nämnda trädformiga svamp via positionsspecifik homologisk rekombination. 10
12. Trädformig svamp enligt nagot av patentkraven 9 - 11, kännetecknad därav, att svampen är Aspergillus, Trichoderma eller PeniciIlium. 15
13. Förfarande för att erhälla en selektionsmarkörgenfri rekombinant trädformig svamp, kännetecknat därav, att det omfattar följande steg: a) ett önskat DNA-fragment och en dominerande och 20 dubbelriktad selektionsmarkörgen integreras i genomet av en trädformig svamp, b) transformanter selekteras, c) selektionsmarkörgenen avslägnas med rekombination mellan , i selektionsmarkörgenens flankerande DNA-upprepningssekvenser, V 25 d) kontraselekteras, som är baserad pä avsaknad av en :· selektionsmarkörgen.
14. Förfarande enligt patentkravet 13, kännetecknat därav, att en sekvens klonas till 5'- eller 3'-sidan av 30 selektionsmarkörgenen, vilken sekvens bildar en upprepningssekvens med en sekvens, som ligger pä 3'- eller ’ 5'-sidan av sekvensen, som avslägnas frän genomet.
: 15. Förfarande enligt patentkravet 13 eller 14, kännetecknat 35 därav, att det önskade DNA-fragmentet omfattar nägot av följande genetiska element eller deras kombinationer: en gen, en cDNA, en promotor, en terminator, ett regleringselement, ' · en intron, en igenkänningssekvens för ett DNA-bindande / 106 „ „ , 114481 protein, ett translationsinitieringsställe eller ett restriktionsställe.
16. Förfarande enligt patentkravet 13 eller 14, kännetecknat 5 därav, att stegen a)-d) repeteras med den erhällna trädformiga svampen med användning av antingen detsamma eller olika önskade DNA-fragment.
17. Förfarande enligt nägot av patentkraven 13 - 16, 10 kännetecknat därav, att selektionsmarkörgenen är en acetamidasgen.
18. Förfarande enligt patentkravet 17, kännetecknat därav, att selektionsmarkörgenen är acetamidasgenen härstammande 15 frän en svamp.
19. Förfarande enligt patentkravet 18, kännetecknat därav, att selektionsmarkörgenen är acetamidasgenen, som är frän arten Aspergillus. 20
20. Förfarande för att framställa en bioaktiv produkt, kännetecknat därav, att förfarandet omfattar odling av en trädformig svamp producerads med ett förfarande enligt nägot : : av patentkraven 13-19. 25
21. Förfarande enligt patentkravet 20, kännetecknat därav, : att den bioaktiva föreningen är ett protein.
22. Förfarande enligt patentkravet 20, kännetecknat därav, 30 att den bioaktiva föreningen är ett antibiotikum.
23. Användning av en trädformig svamp, som har producerats med ett förfarande enligt nägot av patentkraven 13 - 19, för I produktion av en bioaktiv förening. : 35
24. Användning enligt patentkravet 23, kännetecknad därav, ; att den bioaktiva föreningen är ett protein. 114481
25. Användning enligt patentkravet 23, kännetecknad därav, att den bioaktiva föreningen är ett antibiotikum. 1 / » * t
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP93202195 | 1993-07-23 | ||
EP93202195 | 1993-07-23 |
Publications (3)
Publication Number | Publication Date |
---|---|
FI943485A0 FI943485A0 (sv) | 1994-07-22 |
FI943485A FI943485A (sv) | 1995-01-24 |
FI114481B true FI114481B (sv) | 2004-10-29 |
Family
ID=8214006
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
FI943485A FI114481B (sv) | 1993-07-23 | 1994-07-22 | Selektionsmarkörgenfria trådformiga svampstammar, förfarande för deras framställning, och användning av dessa stammar |
FI20041124A FI117388B (sv) | 1993-07-23 | 2004-08-27 | Selektionsmarkörgenfria jäststammar, förfarande för deras framställning och användning av dessa stammar |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
FI20041124A FI117388B (sv) | 1993-07-23 | 2004-08-27 | Selektionsmarkörgenfria jäststammar, förfarande för deras framställning och användning av dessa stammar |
Country Status (13)
Country | Link |
---|---|
US (3) | US5876988A (sv) |
EP (2) | EP1321523A3 (sv) |
JP (2) | JP3833725B2 (sv) |
CN (1) | CN100347301C (sv) |
AT (1) | ATE238425T1 (sv) |
BR (1) | BR9402912A (sv) |
CA (1) | CA2128589C (sv) |
DE (1) | DE69432543T2 (sv) |
DK (1) | DK0635574T3 (sv) |
ES (1) | ES2198410T3 (sv) |
FI (2) | FI114481B (sv) |
IE (1) | IE20030875A1 (sv) |
NZ (2) | NZ299139A (sv) |
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1994
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- 1994-06-30 DK DK94201896T patent/DK0635574T3/da active
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CN1098743A (zh) | 1995-02-15 |
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JPH07147971A (ja) | 1995-06-13 |
FI117388B (sv) | 2006-09-29 |
DK0635574T3 (da) | 2003-08-11 |
IE940591A1 (en) | 1995-01-25 |
CA2128589C (en) | 2005-10-11 |
ATE238425T1 (de) | 2003-05-15 |
EP0635574A1 (en) | 1995-01-25 |
IE20030875A1 (en) | 2004-01-14 |
FI20041124A (sv) | 2004-08-27 |
DE69432543D1 (de) | 2003-05-28 |
FI943485A0 (sv) | 1994-07-22 |
CA2128589A1 (en) | 1995-01-24 |
US5876988A (en) | 1999-03-02 |
NZ264065A (en) | 1998-06-26 |
CN100347301C (zh) | 2007-11-07 |
AU6862894A (en) | 1995-02-02 |
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