FI104639B - Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster - Google Patents
Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster Download PDFInfo
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- FI104639B FI104639B FI892312A FI892312A FI104639B FI 104639 B FI104639 B FI 104639B FI 892312 A FI892312 A FI 892312A FI 892312 A FI892312 A FI 892312A FI 104639 B FI104639 B FI 104639B
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- gene
- pichia pastoris
- isolated
- dna
- aox1
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- 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
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
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- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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- A—HUMAN NECESSITIES
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- C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
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Claims (10)
1. Förfarande för produktion av antigena HBV-partiklar vä-sentligen bestäende av S- och preS2-proteiner ur stammen Pichia pastoris, kännetecknat av att det innefattar ' 5 (a) att transformera stammen Pichia pastoris med en första expressionskassett innehällande en strukturgen för S-protein i hepatitis B-virus, funktionellt förbunden med ett 51-regleromrdde som valts i ett AQXl 5’-reglerom-räde isolerat frän Pichia pastoris eller dhas 5’-reg- 10 leromrädet och en 31 -termineringssekvens av Pichia pas toris; och (b) att transformera stammen Pichia pastoris med en andra expressionskassett innehällande preS2-protein av hepatitis B-virus, funktionellt kopplad tili 5'-reglerom- 15 rädet som valts i ett aoxi 5'-regleromräde isolerat frän Pichia pastoris eller dhas 5'-regleromrädet och en 31-termineringssekvens av Pichia pastoris; och därefter (c) att odla den uppkomna transformerade stammen Pichia pastoris under lämpliga förhällanden för att dstadkomma 20 produktion av nämnda HBV-partiklar.
2. Förfarande enligt patentkrav l, kännetecknat av att nämnda transformation utförs med ätminstone en vektor vald i gruppen bestäende av plasmider eller lineära, integrerande, ·· 25 platsspecifikä vektorer.
3. Förfarande enligt patentkrav 2, kännetecknat av att vektorerna är integrerande. 30
4. Förfarande enligt patentkrav 3, kännetecknat av att man använder • * a) en första integrerande platsspecifik vektor innehällan-de följande seriella sekvens: i) ett första insättningsbart DNA-fragment, 35 ii) en markörgen och ätminstone en expressionskassett inne-hällande en strukturgen för preS2, funktionellt förbunden med ett regleromräde och en 3'-termineringssekvens, och 54 104639 iii) ett andra insättningsbart DNA-fragment i vilket den in-bördes ordningen mellan markörgenen och kassetten för markörgenen (ii) kan växlas; b) och en andra integrerande vektor innehällande följande: 5 i) ätminstone ett insättningsbart DNA-fragment, ii) en markörgen och ätminstone en expressionskassett innehällande en strukturgen för S, funktionellt förbunden med regleromrädet och 3'-termineringssekvensen, i vil-ken den inbördes ordningen mellan markörgenen och kas- 10 setten för komponent (ii) kan växlas.
5. Förfarande enligt patentkrav 4, kännetecknat av att nämnda insättningsbara DNA-fragment erhdlls ur DNA-sekvensen i en gen som isolerats frdn Pichia pastoris och väljs i 15 gruppen bestäende av AOX1, p40, DHAS och HIS4.
6. Förfarande enligt patentkrav 4, kännetecknat av att ät-minstone en expressionskassett innefattar a) ett regleromräde valt i gruppen bestäende av AOXl och
20 DHAS isolerad frän Pichia pastoris, funktionellt för- bundet med b) en strukturgen för preS2 eller S, funktionellt förbunden med c) en 3'-termineringssekvens frän Pichia pastoris vald i ;·· 25 gruppen bestäende av 3 ' - termineringssekvenser som iso lerats frän AOXl-genen, p40-genen, DHAS-genen och HIS4-genen.
7. Förfarande enligt patentkrav 4, kännetecknat av att 30 nämnda markörgen väljs i gruppen bestäende av HIS4 och ARG4, som isolerats frän Pichia pastoris, SUC2 som isolerats frän Saccharomyces cerevisiae och neomycin-gen av Tn903 och TnGOl. 1
8. Förfarande enligt patentkrav 4, kännetecknat av att nämnda vektor innehällande en strukturgen för preS2 innefattar 55 1 0 4 6 3 9 a) ett första insättningsbart DNA-fragment vilket är cirka 1 kb av det 5' AOX1 regleromräde som isolerats frän Pichia pastoris, funktionellt förbundet med b) en strukturgen för preS2, funktionellt förbunden med ' 5 c) 3'-termineringssekvensen för AOX1 som isolerats frän Pichia pastoris, ligerad tili d) en markörgen som är HIS4 som isolerats frän Pichia pastoris, ligerad tili e) ett andra insättningsbart DNA-fragment vilket är cirka 10 0,65 kb av 3' AOX1-termineringssekvensen.
9. Förfarande enligt patentkrav 4, kännetecknat av att nämnda vektor innehällande en strukturgen för S innefattar a) ett insättningsbart DNA-fragment vilket är cirka l kb 15 av 5' AOXl-regleromrädet, som isolerats frän Pichia pastoris, funktionellt förbundet med b) en strukturgen för S, funktionellt förbunden med c) 3' AOX1-termineringssekvensen, som isolerats frän Pichia pastoris, ligerad tili 20 d) en markörgen som är ARG4 som isolerats frän Saccharo-myces cerevisiae, ligerad tili e) ett insättningsbart DNA-fragment vilket är cirka 0,65 kb av 3' A0X1-termineringssekvensen. :·· 25
10. Pichia pastoris PPF1 (NRRL Y-18017) i vilken transforme- rats en kopia av expressionskassetten av pTB05A (jfr exempel II) eller Pichia pastoris PPF1 (NRRL Y-18017), i vilken transformerats tvä kopior av en preS2-gen av pTB05A (jfr exempel II) och en kopia av S-genen. * I
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US19371488A | 1988-05-13 | 1988-05-13 | |
| US19371488 | 1988-05-13 |
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| FI892312A0 FI892312A0 (fi) | 1989-05-12 |
| FI892312A FI892312A (fi) | 1989-11-14 |
| FI104639B true FI104639B (sv) | 2000-03-15 |
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| FI892312A FI104639B (sv) | 1988-05-13 | 1989-05-12 | Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster |
| FI992546A FI107265B (sv) | 1988-05-13 | 1999-11-29 | Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster |
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| Application Number | Title | Priority Date | Filing Date |
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| FI992546A FI107265B (sv) | 1988-05-13 | 1999-11-29 | Expression av hepatitis B, S och PreS2 proteiner i metylotropiska jäster |
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| EP (1) | EP0341746B1 (sv) |
| JP (1) | JP2614314B2 (sv) |
| KR (1) | KR100193701B1 (sv) |
| CN (1) | CN1038668A (sv) |
| AT (1) | ATE151109T1 (sv) |
| AU (1) | AU618596B2 (sv) |
| CA (1) | CA1340617C (sv) |
| DD (1) | DD285373A5 (sv) |
| DE (1) | DE68927921T2 (sv) |
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| FI (2) | FI104639B (sv) |
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| IE (1) | IE81158B1 (sv) |
| IL (1) | IL90161A0 (sv) |
| IN (1) | IN171696B (sv) |
| MX (1) | MX26715A (sv) |
| NO (1) | NO300465B1 (sv) |
| NZ (1) | NZ228948A (sv) |
| PL (1) | PL161997B1 (sv) |
| PT (1) | PT90553A (sv) |
| YU (1) | YU98689A (sv) |
| ZA (1) | ZA893440B (sv) |
Families Citing this family (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5032516A (en) * | 1985-10-25 | 1991-07-16 | Phillips Petroleum Company | Pichia pastoris alcohol oxidase II regulatory region |
| PL168408B1 (pl) * | 1989-08-03 | 1996-02-29 | Smithkline Beecham Biolog | Sposób wytwarzania stransformowanego mikroorganizmu PL |
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| GR76274B (sv) * | 1981-08-04 | 1984-08-04 | Univ California | |
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| US4963483A (en) * | 1987-10-13 | 1990-10-16 | Merck & Co., Inc. | Method for producing hepatitis B virus proteins in yeast |
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1989
- 1989-05-02 IL IL90161A patent/IL90161A0/xx unknown
- 1989-05-02 NZ NZ228948A patent/NZ228948A/en unknown
- 1989-05-09 ZA ZA893440A patent/ZA893440B/xx unknown
- 1989-05-11 KR KR1019890006430A patent/KR100193701B1/ko not_active IP Right Cessation
- 1989-05-11 HU HU892372A patent/HU207532B/hu not_active IP Right Cessation
- 1989-05-11 NO NO891921A patent/NO300465B1/no not_active IP Right Cessation
- 1989-05-12 YU YU98689A patent/YU98689A/sh unknown
- 1989-05-12 AU AU34734/89A patent/AU618596B2/en not_active Expired
- 1989-05-12 DE DE68927921T patent/DE68927921T2/de not_active Expired - Lifetime
- 1989-05-12 FI FI892312A patent/FI104639B/sv active IP Right Grant
- 1989-05-12 PT PT90553A patent/PT90553A/pt not_active Application Discontinuation
- 1989-05-12 PL PL89279426A patent/PL161997B1/pl unknown
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- 1989-05-12 EP EP89108632A patent/EP0341746B1/en not_active Expired - Lifetime
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- 1989-05-12 DK DK235289A patent/DK235289A/da not_active Application Discontinuation
- 1989-05-12 AT AT89108632T patent/ATE151109T1/de not_active IP Right Cessation
- 1989-05-13 CN CN89103330A patent/CN1038668A/zh active Pending
- 1989-05-15 CA CA000599724A patent/CA1340617C/en not_active Expired - Lifetime
- 1989-05-15 JP JP1121313A patent/JP2614314B2/ja not_active Expired - Fee Related
- 1989-05-22 IN IN388/CAL/89A patent/IN171696B/en unknown
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1995
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1997
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1999
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| GB | Transfer or assigment of application |
Owner name: RESEARCH CORPORATION TECHNOLOGIES, INC. |
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| FG | Patent granted |
Owner name: RESEARCH CORPORATION TECHNOLOGIES, INC. |