KR970006505A - 대장균의 복제개시 억제 단백질을 융합 파트너로 이용하는 발현벡터 - Google Patents
대장균의 복제개시 억제 단백질을 융합 파트너로 이용하는 발현벡터 Download PDFInfo
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- KR970006505A KR970006505A KR1019950023571A KR19950023571A KR970006505A KR 970006505 A KR970006505 A KR 970006505A KR 1019950023571 A KR1019950023571 A KR 1019950023571A KR 19950023571 A KR19950023571 A KR 19950023571A KR 970006505 A KR970006505 A KR 970006505A
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- 108090000623 proteins and genes Proteins 0.000 title claims abstract 22
- 102000004169 proteins and genes Human genes 0.000 title claims abstract 16
- 239000013604 expression vector Substances 0.000 title claims abstract 10
- 241000588724 Escherichia coli Species 0.000 title claims abstract 5
- 230000010076 replication Effects 0.000 title claims abstract 5
- 230000004927 fusion Effects 0.000 title claims abstract 4
- 230000000977 initiatory effect Effects 0.000 title claims 2
- 239000012634 fragment Substances 0.000 claims abstract 3
- 108020001507 fusion proteins Proteins 0.000 claims abstract 3
- 102000037865 fusion proteins Human genes 0.000 claims abstract 3
- 230000002401 inhibitory effect Effects 0.000 claims abstract 3
- 238000004519 manufacturing process Methods 0.000 claims abstract 3
- 239000000427 antigen Substances 0.000 claims abstract 2
- 102000036639 antigens Human genes 0.000 claims abstract 2
- 108091007433 antigens Proteins 0.000 claims abstract 2
- 238000000034 method Methods 0.000 claims 3
- 241001288713 Escherichia coli MC1061 Species 0.000 claims 2
- 241000713772 Human immunodeficiency virus 1 Species 0.000 claims 2
- 108091008146 restriction endonucleases Proteins 0.000 claims 2
- 102000055006 Calcitonin Human genes 0.000 claims 1
- 108060001064 Calcitonin Proteins 0.000 claims 1
- 208000031886 HIV Infections Diseases 0.000 claims 1
- 102000002265 Human Growth Hormone Human genes 0.000 claims 1
- 108010000521 Human Growth Hormone Proteins 0.000 claims 1
- 239000000854 Human Growth Hormone Substances 0.000 claims 1
- 101800001690 Transmembrane protein gp41 Proteins 0.000 claims 1
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 claims 1
- 229960004015 calcitonin Drugs 0.000 claims 1
- 238000012258 culturing Methods 0.000 claims 1
- 108091006086 inhibitor proteins Proteins 0.000 claims 1
- 244000005700 microbiome Species 0.000 claims 1
- 239000013598 vector Substances 0.000 claims 1
- 108090000695 Cytokines Proteins 0.000 abstract 1
- 102000004127 Cytokines Human genes 0.000 abstract 1
- SRBFZHDQGSBBOR-HWQSCIPKSA-N L-arabinopyranose Chemical compound O[C@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-HWQSCIPKSA-N 0.000 abstract 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 abstract 1
- 230000004071 biological effect Effects 0.000 abstract 1
- 239000005556 hormone Substances 0.000 abstract 1
- 229940088597 hormone Drugs 0.000 abstract 1
- 239000003910 polypeptide antibiotic agent Substances 0.000 abstract 1
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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
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/70—Vectors or expression systems specially adapted for E. coli
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- 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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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- C07K14/245—Escherichia (G)
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- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/585—Calcitonins
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- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/61—Growth hormone [GH], i.e. somatotropin
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- C07K2319/00—Fusion polypeptide
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- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/16011—Human Immunodeficiency Virus, HIV
- C12N2740/16051—Methods of production or purification of viral material
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- C12N2800/00—Nucleic acids vectors
- C12N2800/80—Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites
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Abstract
본 발명은 대장균에서 대량 발현되는 복제개시 억제 단백질(Ici A) 혹은 그의 단편을 융합 파트너로 이용하는 융합단백질의 발현벡터 및 그를 이용하여 목적 단백질을 제조할 수 있는 방법에 관한 것이다. 본 발명의 발현벡터에 다양한 크기의목적 단백질 유전자를 삽입하여 목적 단백질을 제조할 때, 목적 단백질의 크기에 상관없이 Ici A가 융합된 목적 단백질은 L-arabinose에 의해 정확하게 유도되며, 또한 대량으로 발현됨을 알 수 있었다. 따라서, 본 발명의 융합 파트너로 Ici A 혹은 그의 단편을 삽입하여 제조한 융합단백질의 발현벡터는 다양한 크기의 목적 단백질을 대량으로 제조하고자 하는 경우에 매우 효율적으로 이용될 수 있을 것이다. 특히, 펩타이드 항생제 및 인체호르몬, 각종 싸이토카인 및 항원 등과 같이 생물학적 활성을 갖는 다양한 단백질의 대량생산에 이용될 수 있을 것이다.
Description
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
Claims (9)
- 대장균이 복제개시 억제 단백질 (Ici A) 혹은 그의 단편을 융합 파트너로 이용하는 융합단백질의 발현벡터.
- 대장균의 복제개시 억제 단백질(Ici A) 유전자의 500bp에 해당하는 부분에 SmaI 제한효소 부위가 위치하도록 제조된 Ici A 유전자를 L-arabinose 유도성 발현벡터에 삽입하여 제조한 발현벡터 ΔpMA5S.
- 제2항의 발현벡터 ΔpM5S로 형질전환된 대장균 MC1061:ΔpMA5S(KCCM-10072).
- 대장균의 복제개시 억제 단백질(Ici A) 유전자의 300bp에 해당하는 부분에 SmaI 제한효소 부위가 위치하도록 제조된 Ici A 유전자를 L-arabinose 유도성 발현벡터에 삽입하여 제조한 발현벡터 ΔpMA3S.
- 제4항의 발현벡터 ΔpM3S로 형질전환된 대장균 MC1061:ΔpMA3S(KCCM-10069).
- 제2항 또는 제4항의 벡터에 목적 단백질의 유전자를 도입시킨 발현벡터로 형질전환된 미생물을 배양하여목적 단백질을 수득하는 공정을 포함하는 목적 단백질의 제조방법.
- 제6항에 있어서, 목적 단백질은 칼시토닌(calcitionin)인 것을 특징으로 하는 방법.
- 제6항에 있어서, 목적 단백질은 HIV-1(human immunodeficiency virus type 1)의 gp41 항원인 것을 특징으로 하는 방법.
- 제6항에 있어서, 목적 단백질은 인간 성장 호르몬인 것을 특징으로 하는 방법.※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1019950023571A KR0150338B1 (ko) | 1995-07-31 | 1995-07-31 | 대장균의 복제개시 억제 단백질을 융합 파트너로 이용하는 발현벡터 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1019950023571A KR0150338B1 (ko) | 1995-07-31 | 1995-07-31 | 대장균의 복제개시 억제 단백질을 융합 파트너로 이용하는 발현벡터 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR970006505A true KR970006505A (ko) | 1997-02-21 |
| KR0150338B1 KR0150338B1 (ko) | 1998-08-17 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1019950023571A KR0150338B1 (ko) | 1995-07-31 | 1995-07-31 | 대장균의 복제개시 억제 단백질을 융합 파트너로 이용하는 발현벡터 |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR0150338B1 (ko) |
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1995
- 1995-07-31 KR KR1019950023571A patent/KR0150338B1/ko not_active IP Right Cessation
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| Publication number | Publication date |
|---|---|
| KR0150338B1 (ko) | 1998-08-17 |
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