KR20040084520A - Ｃｒｅ-ＬｏｘＰ 시스템을 이용한 유전자 트랩의 가역적조절방법 및 트랩벡터 - Google Patents
Ｃｒｅ-ＬｏｘＰ 시스템을 이용한 유전자 트랩의 가역적조절방법 및 트랩벡터 Download PDFInfo
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- KR20040084520A KR20040084520A KR1020030019658A KR20030019658A KR20040084520A KR 20040084520 A KR20040084520 A KR 20040084520A KR 1020030019658 A KR1020030019658 A KR 1020030019658A KR 20030019658 A KR20030019658 A KR 20030019658A KR 20040084520 A KR20040084520 A KR 20040084520A
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- loxp
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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
- C12N15/635—Externally inducible repressor mediated regulation of gene expression, e.g. tetR inducible by tetracyline
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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
- C12N15/65—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression using markers
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- C—CHEMISTRY; METALLURGY
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- 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
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
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Abstract
Description
Claims (12)
- 스플라이싱 수용체(splicing acceptor; SA), IRES(internal ribosomal entry site) 및 외부 유전자를 포함하는 트랩 벡터를 게놈 DNA의 인트론 부위에 삽입시켜 외부 유전자를 발현시키는 유전자 트랩의 조절방법에 있어서,상기 트랩벡터는 LoxP 및 LoxP 역방향체 사이에 SA 및 IRES을 포함하며, 외재적인 Cre 재조합효소에 의하여 트랩벡터내 SA 및 IRES의 방향성이 조절되는 것을 포함하는 유전자 트랩의 조절방법.
- 제 1항에 있어서, 상기 트랩벡터는 외부 유전자 상부에 5'-LoxP-SERI-AS-LoxP 역방향체-3' 순으로 배열된 염기서열을 포함하여 Cre 재조합효소 존재시 5'-LoxP-SA-IRES-LoxP 역방향체-3'순으로 SA 및 IRES 염기서열이 전위되어 외부유전자가 발현되는 것인 유전자 트랩의 조절방법.
- 제 1항에 있어서, 상기 트랩벡터는 외부 유전자 상부에 5'-LoxP-SA-IRES-LoxP 역방향체-3' 순으로 배열된 염기서열을 포함하여 Cre 재조합효소 존재시 5'-LoxP-SERI-AS-LoxP 역방향체-3'순으로 SA 및 IRES 염기서열이 전위되어 외부유전자의 발현이 저해되는 것인 유전자 트랩의 조절방법.
- 제 1항에 있어서, 상기 트랩벡터는 PGK-puro-loxP 카세트 벡터 또는 PGK-neo-loxP 카세트 벡터인 것인 유전자 트랩의 조절방법.
- (a) LoxP, SA, IRES, LoxP 역방향체 및 외부 유전자를 포함하며, LoxP와 LoxP 역방향체 사이의 염기서열은 Cre 재조합효소에 의하여 역전위되는 트랩벡터를 도입하여 동물배를 재구성하는 단계 및;(b) 상기 재구성된 동물 배로부터 동물을 발생시키는 단계를 포함하는 유전자 이식동물 제조방법.
- 제 5항에 있어서, 상기 (b) 단계에서 발생된 동물을 Cre 유전자 이식 동물과 교배시켜 외부유전자를 발현하는 동물을 수득하는 단계를 더욱 포함하는 것인 유전자 이식동물 제조방법.
- 제 5항 또는 6항의 방법으로 제조된 유전자 이식동물.
- LoxP 및 LoxP 역방향체 사이에 위치한 SA 및 IRES, 외부유전자 및 선별마커를 포함하는 트랩벡터.
- 제 8항에 있어서, 상기 트랩벡터는 동물 게놈 DNA의 인트론상에 삽입되고, Cre 재조합효소에 의하여 SA 및 IRES 염기서열이 역위가능하도록 포함하는 것인 트랩벡터.
- 제 8항에 있어서, 상기 트랩벡터는 동물 게놈 DNA의 인트로상에 삽입된 상태에서 Cre 재조합효소에 의하여 5'-SA-IRES-3' 순으로 배열된 염기서열이 3'-SERI-AS-5'순으로 역위되어 외부유전자를 동물 게놈 DNA의 프로모터에 의하여 발현시키는 것인 트랩벡터.
- 제 8항 내지 10항 중 어느 한항에 따른 트랩벡터가 형질도입된 형질전환세포.
- 제 11항에 있어서, 상기 형질전환세포는 배아간세포, 간세포 및 수정난으로 이루어진 군으로부터 선택된 것인 형질전환세포.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020030019658A KR100547164B1 (ko) | 2003-03-28 | 2003-03-28 | Ｃｒｅ-ＬｏｘＰ 시스템을 이용한 유전자 트랩의 가역적 조절방법 및 트랩벡터 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020030019658A KR100547164B1 (ko) | 2003-03-28 | 2003-03-28 | Ｃｒｅ-ＬｏｘＰ 시스템을 이용한 유전자 트랩의 가역적 조절방법 및 트랩벡터 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20040084520A true KR20040084520A (ko) | 2004-10-06 |
| KR100547164B1 KR100547164B1 (ko) | 2006-01-26 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020030019658A Expired - Lifetime KR100547164B1 (ko) | 2003-03-28 | 2003-03-28 | Ｃｒｅ-ＬｏｘＰ 시스템을 이용한 유전자 트랩의 가역적 조절방법 및 트랩벡터 |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR100547164B1 (ko) |
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- 2003-03-28 KR KR1020030019658A patent/KR100547164B1/ko not_active Expired - Lifetime
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| Publication number | Publication date |
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| KR100547164B1 (ko) | 2006-01-26 |
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