KR950025103A - New Biscistronic Vector - Google Patents

New Biscistronic Vector Download PDF

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KR950025103A
KR950025103A KR1019940002967A KR19940002967A KR950025103A KR 950025103 A KR950025103 A KR 950025103A KR 1019940002967 A KR1019940002967 A KR 1019940002967A KR 19940002967 A KR19940002967 A KR 19940002967A KR 950025103 A KR950025103 A KR 950025103A
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South Korea
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gene
vector
expression vector
promoter
selection
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KR1019940002967A
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KR0128874B1 (en
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조중명
류왕식
양재영
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성재갑
엘지화학 주식회사
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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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/505—Erythropoietin [EPO]
    • 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
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/16011—Herpesviridae
    • C12N2710/16111—Cytomegalovirus, e.g. human herpesvirus 5
    • C12N2710/16141—Use of virus, viral particle or viral elements as a vector

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
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  • General Health & Medical Sciences (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
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  • Gastroenterology & Hepatology (AREA)
  • Physics & Mathematics (AREA)
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  • Virology (AREA)
  • Plant Pathology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
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  • Medicinal Chemistry (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

본 발명은 외래유전자와 선별유전자의 코딩하는 2개의 오픈리딩 프레임을 갖는 mRNA가 한 개의 프로모로부터 전사되도록 하는 동물 발현용 바이시스트로닉 발현벡터에 관한 것이다.The present invention relates to a biscistronic expression vector for animal expression, which allows mRNA having two open reading frames encoding a foreign gene and a selection gene to be transcribed from one promo.

Description

신규 바이시스트로닉 벡터New Biscistronic Vector

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음Since this is an open matter, no full text was included.

제1조는 동물 세포 발현용 바이시스트로닉(bicistronic) 벡터인 pCED2의 클로닝 과정을 도식화한 것이고,Article 1 is a schematic of the cloning process of pCED2, a bicistronic vector for animal cell expression,

제3도는 클로닝 과정에서 사용된 천연 리더 서열을 갖고 있는 인간 적혈구 성장 촉진인자 유전자의 염기 서열을 나타낸 것이고,Figure 3 shows the nucleotide sequence of the human erythrocyte growth promoter gene having a natural leader sequence used in the cloning process,

제4도는 본 발명의 pCED2 벡터에 적혈구 성장 촉진인자(erythropoietin,EPO)의 유전자를 삽입하여 포유 동물에서 발현을 시킨 후 웨스턴 블럿팅한 결과이다.4 is a result of Western blotting after expression in a mammal by inserting a gene of erythropoietin (EPO) into the pCED2 vector of the present invention.

Claims (12)

하나의 프로모터 조절하에 선별 유전자와 외래 유전자가 발현되는 동물 세포발현용 바이시스트로닉 벡터.Biscistronic vectors for animal cell expression in which select and foreign genes are expressed under one promoter. 제1항에 있어서, 상기 선별유전자가 다이하드로폴레이트 리덕테이즈 유전자인 벡터.The vector of claim 1, wherein the selection gene is a dihydrofolate reductase gene. 제2항에 있어서, 다이하드로 폴레이트 리덕테이즈 유전자 사이에 엔세팔로 미오카디티스 바이러스(Encephalpmyocarditis virus)의 리더서열이 삽입되고, 상기 리더서열과 프로모터 사이에 외래유전자가 치하는 벡터.The vector according to claim 2, wherein a leader sequence of Encephalpmyocarditis virus is inserted between the diehard folate reductase genes, and a foreign gene is inserted between the leader sequence and the promoter. 제1항에 있어서, 상기 프로모터가 사이토메갈로 바이러스 초기 프로모터인 벡터.The vector of claim 1, wherein said promoter is a cytomegalovirus initial promoter. 제4항에 있어서, 발현벡터 pCED2의 다중 클로닝 부위에 외부 유전자가 삽입된 벡터.The vector of claim 4, wherein an external gene is inserted into the multiple cloning site of the expression vector pCED2. 제5항에 있어서, 상기외래 유전자가 적혈구 성장촉진인자(EPO) 유전자인 발현벡터 pCED2 L-EPO.The expression vector pCED2 L-EPO according to claim 5, wherein the foreign gene is an erythrocyte growth promoting factor (EPO) gene. 제1항내지 제6항중 어느 한 항에 발현벡터를 포함하는 숙주세포.A host cell comprising the expression vector of claim 1. 제7항에 있어서, 상기 발현벡터에 의해 형질감염된 동물세포.The animal cell of claim 7, wherein the animal cell is transfected with the expression vector. 제8항의 동물세포를 배양함을 포함하는, 상기 발현벡터에 삽입된 외래 유전자에 코드화된 단백질을 제조하는 방법.A method for preparing a protein encoded in a foreign gene inserted into the expression vector, comprising culturing the animal cell of claim 8. 제9항에 있어서, 발현벡터 pCED2 L-EPO로 형질감염된 동물 세포를 배양함을 포함하는 적혈구성장 촉진 인자의 제조방법.10. The method of claim 9, comprising culturing animal cells transfected with the expression vector pCED2 L-EPO. 제9항에 있어서, 배양시 상기 발현벡터에 포함된 선별유전자를 선별할 수 있는 선별 약품을 첨가하는 방법.10. The method of claim 9, wherein a selection agent capable of selecting a selection gene included in the expression vector is added during the culture. 제11항에 있어서, 상기 선별유전자가 다이하드로폴레이트 리덕테이즈 유전자인 경우 선별약품으로 메소트랙세이트를 첨가하는 방법.12. The method according to claim 11, wherein the mesotraxate is added to the selection agent when the selection gene is a dihydrofolate reductase gene. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019940002967A 1994-02-19 1994-02-19 A novel bicistronic vector KR0128874B1 (en)

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Application Number Priority Date Filing Date Title
KR1019940002967A KR0128874B1 (en) 1994-02-19 1994-02-19 A novel bicistronic vector

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KR950025103A true KR950025103A (en) 1995-09-15
KR0128874B1 KR0128874B1 (en) 1998-04-04

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