KR970070207A - Expression vector containing promoter of elongation factor 2 (EF-2) - Google Patents

Expression vector containing promoter of elongation factor 2 (EF-2) Download PDF

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KR970070207A
KR970070207A KR1019970038369A KR19970038369A KR970070207A KR 970070207 A KR970070207 A KR 970070207A KR 1019970038369 A KR1019970038369 A KR 1019970038369A KR 19970038369 A KR19970038369 A KR 19970038369A KR 970070207 A KR970070207 A KR 970070207A
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promoter
expression vector
elongation factor
present
various
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KR1019970038369A
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KR100222919B1 (en
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강윤세
정은주
하영술
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김충원
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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

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Abstract

본 발명은 연장인자 2(elongation factor 2, EF-2)의 프로모터(promoter)를 포함하는 발현벡터에 관한 것이다. 좀 더 구체적으로, 본 발명은 닭의 EF-2 프로모터 약 650bp의 염기서열, 및 생리학적으로 중요한 각종 진핵세포 단백질을 효율적으로 대량 발현시키기 위하여 전기 염기서열의 유전자를 프로모터로 이용하는 발현벡터에 관한 것이다. EF-2프로모터를 포함하는 본 발명의 발현벡터는 폭넓은 숙주세포에서 목적단백질을 대량으로 발현할 수 있으므로, 고가의 생리활성물질 등을 보다 효율적이고도 경제적으로 제조할 수 있으며, 또한 SV-40 및 EF-1α프로모터와는 달리 혈청이 없는 배지에서도 다양한 숙주세포에서 그 프로모터의 활성을 가질 수 있으므로, 산업적인 측면에서 훨씬 경제적이라고 할 수 있다.The present invention relates to an expression vector comprising a promoter of elongation factor 2 (EF-2). More specifically, the present invention relates to a nucleotide sequence of about 650 bp of EF-2 promoter in chickens and an expression vector using genes of the above nucleotide sequences as promoters for efficient mass expression of various physiologically important eukaryotic proteins. . Since the expression vector of the present invention including the EF-2 promoter can express a large amount of the protein of interest in a wide range of host cells, it is possible to more efficiently and economically prepare expensive bioactive substances and the like. Unlike the EF-1α promoter, even in a medium without serum, the promoter may have activity of various promoters in various host cells, and thus, may be more economically industrially.

Description

연장인자 2(EF-2)의 프로모터를 포함하는 발현벡터Expression vector containing promoter of elongation factor 2 (EF-2)

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음As this is a public information case, the full text was not included.

제1도는 닭의 연장인자2(elongation factor 2 EF-2) 프로모터 약 650bp의 염기서열을 나타낸다. 제2도는 닭의 EF-2 프로모터 유전자를 이용하여 진핵세포에서의 발현 벡터인 pEF2-CAT 운반벡터를 구축하는 모식도이다. 제3도는 NIH3T3 숙주세포에 각 운반벡터를 도입하고 FCS(fetal calf serum)가 없는 조건에서 배양한 경우의 CAT(chloramphenicol acetyltransferase) 효소활성을 나타내는 TLC 사진이다.Figure 1 shows a nucleotide sequence of about 650 bp of elongation factor 2 (EF-2) promoter in chickens. 2 is a schematic diagram of constructing a pEF2-CAT transport vector, which is an expression vector in eukaryotic cells, using chicken EF-2 promoter gene. FIG. 3 is a TLC photograph showing the CAT (chloramphenicol acetyltransferase) enzyme activity when each transport vector is introduced into NIH3T3 host cells and cultured in the absence of fetal calf serum (FCS).

Claims (3)

하기와 같은 염기서열을 갖는 연장인자 2(elongation factor 2, EF-2)의 프로모터 :Promoter of elongation factor 2 (EF-2) having the following nucleotide sequence: 1 CTGCAGGTCGACTAGAGTCGACCCCTCTTTGCTCGCCCATTGTGGXXXXT1 CTGCAGGTCGACTAGAGTCGACCCCTCTTTGCTCGCCCATTGTGGXXXXT 51 TCGCTGCTAGTTTCCTCCTCCTCGTCAGCAATATCAGGACACCACGGCTC51 TCGCTGCTAGTTTCCTCCTCCTCGTCAGCAATATCAGGACACCACGGCTC 101 CATTTCCATGCGCAGCTGGAGCGGAGACGCCGGGAGGGGCGGCGNTTGCC101 CATTTCCATGCGCAGCTGGAGCGGAGACGCCGGGAGGGGCGGCGNTTGCC 151 CACGCCANGGNACAGTCCNGNTGCGGCCCGGGCCGCGCCCCGCNGTATCC151 CACGCCANGGNACAGTCCNGNTGCGGCCCGGGCCGCGCCCCGCNGTATCC 201 GACACAGCGCTCCGGGCGCAGGCGGAGGCGCTCTCGTGGGCGCGGGGAGC201 GACACAGCGCTCCGGGCGCAGGCGGAGGCGCTCTCGTGGGCGCGGGGAGC 251 GCCCCGAATACGTCCTCCGGGTCGCGCCCGCCGTGGCNGAGTTGTTTCCG251 GCCCCGAATACGTCCTCCGGGTCGCGCCCGCCGTGGCNGAGTTGTTTCCG 301 CTGCGGGTTCTGAGCGGGGACTGCGTGGAGCCCGACTGCGGCCCTTGGAG301 CTGCGGGTTCTGAGCGGGGACTGCGTGGAGCCCGACTGCGGCCCTTGGAG 351 GACCTCACCGCCCGACCATAGAGCTGCCGGCAGCCGGGTGCTAGAGAGGC351 GACCTCACCGCCCGACCATAGAGCTGCCGGCAGCCGGGTGCTAGAGAGGC 401 GCATCTGCGCACGCGCACGCGCAGGCTCCTCCCCCCGGCCGGCGCTGCCC401 GCATCTGCGCACGCGCACGCGCAGGCTCCTCCCCCCGGCCGGCGCTGCCC 451 TATAAAAGGGGGGCGGTTGACGTCAGCGTTCTCTTCCGCCCGCTCTGCGC451 TATAAAAGGGGGGCGGTTGACGTCAGCGTTCTCTTCCGCCCGCTCTGCGC 501 TCGCCAGCGAGACAGGGCCGACGCCGCCGCCATTGCTGTACCCCCCCGCC501 TCGCCAGCGAGACAGGGCCGACGCCGCCGCCATTGCTGTACCCCCCCGCC 551 CCGGAGAATCCGCCTCCATCCGCCACC551 CCGGAGAATCCGCCTCCATCCGCCACC 상기에서, X는 결정되지 않은 아미노산 잔기 20개를 나타내고, N은 결정되지 않은 아미노산 잔기 1개를 나타낸다.In the above, X represents 20 undetermined amino acid residues and N represents 1 undetermined amino acid residues. 제1항의 연장인자 2(elongation factor, EF-2) 프로모터를 목적 단백질의 발현을 위한 프로모터로서 이용하는 발현벡터.An expression vector using the elongation factor (EF-2) promoter of claim 1 as a promoter for expression of a target protein. 제1항의 연장인자 2(elongation factor 2, EF-2) 프로모터를 포함하는 운반벡터 pEF2-CAT(KFCC-10981).A carrier vector pEF2-CAT (KFCC-10981) comprising the elongation factor 2 (EF-2) promoter of claim 1. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019970038369A 1997-08-12 1997-08-12 Novel expression vector which includes elongation factor 2(ef-2) KR100222919B1 (en)

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KR100222919B1 KR100222919B1 (en) 1999-10-01

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