KR960010864A - 동물 세포의 형질 감염을 위한 재조합 dna 바이러스 벡터 - Google Patents
동물 세포의 형질 감염을 위한 재조합 dna 바이러스 벡터 Download PDFInfo
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- KR960010864A KR960010864A KR1019950029877A KR19950029877A KR960010864A KR 960010864 A KR960010864 A KR 960010864A KR 1019950029877 A KR1019950029877 A KR 1019950029877A KR 19950029877 A KR19950029877 A KR 19950029877A KR 960010864 A KR960010864 A KR 960010864A
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- 108020004511 Recombinant DNA Proteins 0.000 title claims abstract 16
- 239000013598 vector Substances 0.000 title claims abstract 14
- 210000004102 animal cell Anatomy 0.000 title claims abstract 12
- 241000700605 Viruses Species 0.000 title claims abstract 10
- 238000001890 transfection Methods 0.000 title claims 3
- 108020004414 DNA Proteins 0.000 claims abstract description 11
- 108091034057 RNA (poly(A)) Proteins 0.000 claims abstract 17
- 108090000623 proteins and genes Proteins 0.000 claims abstract 15
- 239000013603 viral vector Substances 0.000 claims abstract 11
- 230000010076 replication Effects 0.000 claims abstract 10
- 108010091086 Recombinases Proteins 0.000 claims abstract 5
- 102000053602 DNA Human genes 0.000 claims abstract 3
- 108020005091 Replication Origin Proteins 0.000 claims abstract 3
- 239000012634 fragment Substances 0.000 claims abstract 3
- 238000000034 method Methods 0.000 claims 5
- 241000701161 unidentified adenovirus Species 0.000 claims 4
- 241000701022 Cytomegalovirus Species 0.000 claims 2
- 101000834253 Gallus gallus Actin, cytoplasmic 1 Proteins 0.000 claims 2
- 102100021519 Hemoglobin subunit beta Human genes 0.000 claims 2
- 108091005904 Hemoglobin subunit beta Proteins 0.000 claims 2
- 241000283973 Oryctolagus cuniculus Species 0.000 claims 2
- 101710200251 Recombinase cre Proteins 0.000 claims 2
- 239000000370 acceptor Substances 0.000 claims 2
- 239000003623 enhancer Substances 0.000 claims 2
- 230000002463 transducing effect Effects 0.000 claims 2
- 238000011144 upstream manufacturing Methods 0.000 claims 2
- 241000588724 Escherichia coli Species 0.000 claims 1
- 108091028043 Nucleic acid sequence Proteins 0.000 claims 1
- 241000125945 Protoparvovirus Species 0.000 claims 1
- 241000700647 Variola virus Species 0.000 claims 1
- 210000004027 cell Anatomy 0.000 claims 1
- 125000004122 cyclic group Chemical group 0.000 claims 1
- 238000001415 gene therapy Methods 0.000 claims 1
- 230000006798 recombination Effects 0.000 claims 1
- 238000005215 recombination Methods 0.000 claims 1
- 239000000758 substrate Substances 0.000 claims 1
- 238000003151 transfection method Methods 0.000 claims 1
- 241001529453 unidentified herpesvirus Species 0.000 claims 1
- 208000026350 Inborn Genetic disease Diseases 0.000 abstract 1
- 102000018120 Recombinases Human genes 0.000 abstract 1
- 230000007547 defect Effects 0.000 abstract 1
- 208000016361 genetic disease Diseases 0.000 abstract 1
- 238000002105 Southern blotting Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 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
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
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- C12N2710/10011—Adenoviridae
- C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
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- C12N2800/00—Nucleic acids vectors
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Abstract
프로모터, 리컴비나제 유전자 및 폴리 (A) 서열을 포함하는 재조합 DNA 바이러스 벡터, 및 두개의 리컴비나제-인식 서열과 그 외에 상기 동물 세포에서 작용할 수 있는 복제 기원, 프로모터, 외부 유전자 및 폴리 (A) 서열을 더 포함하며 그 복제 기원, 프로모터, 외부 유전자 및 폴리 (A) 서열을 모두는 두 리컴비나제-인식 서열 사이에 위치하는 재조합 DNA 바이러스 벡터 모두로 동물 세포를 공-형질 감염시킨다. 그런 후, 공-형질 감염된 동물 세포에서, 복제 기원, 프로모터 외부 유전자 및 폴리 (A) 서열을 포함하는 DNA 절편을 또 다른 벡터에서 발현된 리컴비나제의 작용에 의하여 그 벡터로 부터 절단한다. 그 DNA 절편은 복제 기원에 의하여 공-형질 감염된 동물 세포에서 자가적으로 복제할 수 있는 환상의 DNA 분자를 형성하여, 외부 유전자는 계속해서 발현한다.따라서,환자에세 결함이 있는 유전자를 외부 유전자로 사용하는 상기 두DNA 바이러스 벡터의 조합은 유전적 질병을 가진 환자의 치료에 특별히 유용하다
Description
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 본 발명의 형질 도입으로 수득된 DNA를 블러팅(Southern blotting)한 도면이다.
Claims (15)
- 프로모터, 리컴비나제 유전자 및 폴리 (A) 서열을 포함하는 동물 세포의 형질 감염용 재조합 DNA 바이러스 벡터.
- 제1항에 있어서, 상기 DNA 바이러스 벡터가 아데노바이러스 벡터임을 특징으로 하는 재조합 DNA 바이러스 벡터.
- 제2항에 있어서, 상기 리컴비나제 유전자 이.콜리 P1파지로 부터 유래하는 리컴비나제 Cre 유전자임을 특징으로 하는 재조합 DNA 바이러스 벡터.
- 제1항 내지 3항 중 어느 한 항에 있어서, 상기 프로모터 및 폴리 (A) 서열은 사이토메갈로바이러스 인핸서(enhancer), 닭의 β-액틴 프로모터, 및 토끼 β-글로빈 스플라이싱(splicing) 억셉터 및 폴리 (A) 서열을 포함하는 히브리드 포로모터에 포함됨을 특징으로 하는 재조합 DNA 바이러스 벡터.
- 두 개의 리컴비나제-인식 서열, 동물 세포에서 작용할 수 있는 복제 기원, 프로모터, 외부 유전자 및 폴리 (A) 서열을 함유하며, 상기 복제기원, 프로모터, 외부 유전자 및 폴리 (A) 서열 모두가 두 리컴비나제-인식 서열의 사이에 위치하는 상기 동물 세포의 형질 감염용 재조합 DNA 바이러스 벡터.
- 제5항에 있어서, 상기 DNA 바이러스 벡터가 아데노바이러스 벡터임을 특징으로 하는 재조합 DNA 바이러스 벡터.
- 제6항에 있어서, 두 리컴비나제-인식 서열중의 하나의 상류(upstream) 쪽에서 상기 복제 기원, 프로모터, 외부 유전자 및 폴리 (A) 서열이 그 순서대로 위치함을 특징으로 하는 재조합 DNA 바이러스 벡터.
- 제6항에 있어서, 두 리컴비나제-인식 서열중의 하나의 상류 쪽에서 외부 유전자, 폴리 (A) 서열, 복제 기원 및 프로모터가 그 순서대로 위치함을 특징으로 하는 재조합 DNA 바이러스 벡터.
- 제5항 내지 8항 중 어느 한 항에 있어서, 상기 리컴비나제-인식 서열이 리컴비나제 Cre의 기질인 loxP를 코딩하는 DNA 서열 임을 특징으로 하는 재조합 DNA 바이러스 벡터.
- 제5항 내지 8항 중 어느 한 항에 있어서, 상기 복제 기원이 바이러스 또는 동물 세포로 부터 유래함을 특징으로 하는 재조합 DNA 바이러스 벡터.
- 제10항에 있어서, 상기 복제 기원이 파포바바이러스, 헤르페스바이러스, 아데노바이러스, 천연두 바이러스 및 파르보바이러스로 부터 유래하는 복제 기원의 군으로 부터 선택됨을 특징으로 하는 재조합 DNA 바이러스 벡터.
- 제5항 내지 8항 중 어느 한 항에 있어서, 상기 프로모터 및 폴리 (A) 서열은 사이토메갈로바이러스 인핸서(enhancer), 닭의 β-액틴 프로모터, 및 토끼β-글로빈 스프라이싱(splicing) 억셉터 및 폴리 (A) 서열을 포함하는 히브리드 프로모터에 포함됨을 특징으로 하는 재조합 DNA 바이러스 벡터.
- 하기 단계를 특징으로 하는 외부 유전자를 동물 세포로 형질 도입하는 방법 : 프로모터, 리컴비나제 유전자 및 폴리 (A) 서열을 포함하는 재조합 DNA 바이러스 벡터, 및 두개의 리컴비나제-인식 서열, 동물 세포에서 작용할 수 있는 복제 기원, 프로모터, 외부 유전자 및 폴리 (A) 서열을 포함하는 그 복제 기원, 프로모터, 외부 유전자 및 폴리 (A) 서열 모두는 두 리컴비나제-인식 서열 사이에 위치하는 재조합 DNA 바이러스벡터 모두를, 상기 동물 세포로 공-형질 감염시키고; 상기 복제 기원, 프로모터, 외부 유전자 및 폴리 (A) 서열을 함유하는 DNA 절편을 절단하여 환상의 DNA 분자를 만들고; 및 상기 환상의 DNA 분자를 공-형질 감염된 동물 세포내에서 자가 복제를 수행한다.
- 제13항에 있어서, 상기 두 DNA 바이러스 벡터가 각각 아데노바이러스 벡터임을 특징으로 하는 외부 유전자를 동물 세포로 형질 도입하는 방법.
- 제13항 또는 14항에 따른 형질 감염 방법을 유전자 치료에 사용하는 것을 특징으로 하는, 세포로 인체 유전자를 형질 도입하는 방법.※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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JP25131294A JP4216350B2 (ja) | 1994-09-19 | 1994-09-19 | 動物細胞感染用の組換えdnaウイルスベクター |
JP94-251312 | 1994-09-19 |
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KR960010864A true KR960010864A (ko) | 1996-04-20 |
KR100379654B1 KR100379654B1 (ko) | 2004-12-03 |
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KR1019950029877A KR100379654B1 (ko) | 1994-09-19 | 1995-09-13 | 동물세포의형질감염을위한재조합dna바이러스벡터 |
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US (1) | US5817492A (ko) |
EP (1) | EP0704534B1 (ko) |
JP (1) | JP4216350B2 (ko) |
KR (1) | KR100379654B1 (ko) |
CN (1) | CN1077919C (ko) |
AT (1) | ATE321874T1 (ko) |
AU (1) | AU698250B2 (ko) |
CA (1) | CA2157063C (ko) |
DE (1) | DE69534902T2 (ko) |
NZ (1) | NZ272883A (ko) |
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DE19530412A1 (de) * | 1995-08-18 | 1997-02-20 | Harald Von Prof Dr Melchner | Selbst deletierende retrovirale Vektoren für die Gentherapie |
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FR2749857B1 (fr) * | 1996-06-12 | 1998-08-14 | Centre Nat Rech Scient | Generation de molecules replicatives in vivo |
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Publication number | Priority date | Publication date | Assignee | Title |
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CA1293460C (en) * | 1985-10-07 | 1991-12-24 | Brian Lee Sauer | Site-specific recombination of dna in yeast |
NZ225470A (en) * | 1987-07-21 | 1990-06-26 | Du Pont | Stable and viable recombinant viral vectors |
JP2824434B2 (ja) * | 1989-11-28 | 1998-11-11 | 財団法人化学及血清療法研究所 | 新規発現ベクター |
GB9123929D0 (en) * | 1991-11-11 | 1992-01-02 | Wellcome Found | Novel expression system |
US5919676A (en) * | 1993-06-24 | 1999-07-06 | Advec, Inc. | Adenoviral vector system comprising Cre-loxP recombination |
JP3770333B2 (ja) * | 1995-03-15 | 2006-04-26 | 大日本住友製薬株式会社 | 組換えdnaウイルスおよびその製造方法 |
FR2737501B1 (fr) * | 1995-07-31 | 1997-10-24 | Transgene Sa | Nouveaux virus auxiliaires pour la preparation de vecteurs viraux recombinants |
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1994
- 1994-09-19 JP JP25131294A patent/JP4216350B2/ja not_active Expired - Lifetime
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- 1995-08-25 AU AU30248/95A patent/AU698250B2/en not_active Expired
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- 1995-09-13 KR KR1019950029877A patent/KR100379654B1/ko not_active IP Right Cessation
- 1995-09-14 EP EP95306498A patent/EP0704534B1/en not_active Expired - Lifetime
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JPH0884589A (ja) | 1996-04-02 |
CN1077919C (zh) | 2002-01-16 |
US5817492A (en) | 1998-10-06 |
NZ272883A (en) | 1996-11-26 |
CN1132791A (zh) | 1996-10-09 |
JP4216350B2 (ja) | 2009-01-28 |
EP0704534A2 (en) | 1996-04-03 |
EP0704534A3 (en) | 1997-06-11 |
ATE321874T1 (de) | 2006-04-15 |
AU698250B2 (en) | 1998-10-29 |
DE69534902D1 (de) | 2006-05-18 |
AU3024895A (en) | 1996-04-04 |
EP0704534B1 (en) | 2006-03-29 |
DE69534902T2 (de) | 2006-11-09 |
CA2157063C (en) | 2005-11-01 |
CA2157063A1 (en) | 1996-03-20 |
KR100379654B1 (ko) | 2004-12-03 |
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