KR940021575A - Method for producing non-structural 5-1.2 protein of hepatitis C virus - Google Patents

Method for producing non-structural 5-1.2 protein of hepatitis C virus Download PDF

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KR940021575A
KR940021575A KR1019930026611A KR930026611A KR940021575A KR 940021575 A KR940021575 A KR 940021575A KR 1019930026611 A KR1019930026611 A KR 1019930026611A KR 930026611 A KR930026611 A KR 930026611A KR 940021575 A KR940021575 A KR 940021575A
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protein
structural
virus
hepatitis
expression vector
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KR1019930026611A
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KR100285585B1 (en
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조중명
김천형
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최근선
주식회사 럭키
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    • C—CHEMISTRY; METALLURGY
    • 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
    • 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
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/95—Fusion polypeptide containing a motif/fusion for degradation (ubiquitin fusions, PEST sequence)
    • 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
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/24011—Flaviviridae
    • C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
    • C12N2770/24251—Methods of production or purification of viral material

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

Abstract

본 발명은 HCV의 비구조 5-1.2 단백질의 유전자를 유비퀴틴 유전자와 결합된 상태로 포함하여 그의 발현시 HCV의 비구조 5-1.2 단백질과 유비퀴틴의 융합단백질을 생산하는 발현벡터, 상기 발현벡터를 사용하여 HCV의 비구조 5-1.2 단백질을 대량 생산하는 방법 및 이에 의해 생산된 유비퀴틴에 융합된 HCV의 비구조 5-1.2 단백질에 관한 것이다.The present invention uses an expression vector that comprises a non-structural 5-1.2 protein gene of HCV in combination with a ubiquitin gene to produce a fusion protein of non-structural 5-1.2 protein and ubiquitin of HCV, and uses the expression vector. To a method for mass production of the nonstructural 5-1.2 protein of HCV and to the nonstructural 5-1.2 protein of HCV fused to the ubiquitin produced thereby.

Description

씨(C)형 간염 바이러스의 비구조 5-1.2 단백질의 제조방법Method for producing non-structural 5-1.2 protein of hepatitis C virus

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

제 1 도는 C형 간염 바이러스의 비구조 5-1.2 유전자의 염기서열 및 아미노산 서열을 나타낸 것이고Figure 1 shows the nucleotide sequence and amino acid sequence of the nonstructural 5-1.2 gene of hepatitis C virus.

제 2 도는 C형 간염 바이러스의 비구조 5-1.2 유전자의 연결 전략 및 생성된 발현 벡터를 도시한 것이고Figure 2 depicts the linkage strategy of the nonstructural 5-1.2 gene of hepatitis C virus and the resulting expression vector

제 3 도의 A는 비구조 5-1.2 유전자의 발현을 SDS 폴리아크릴아미드젤 전기영동으로 확인한 결과를 나타낸 것이며, 제3도의 B는 비구조 5-1.2 유전자의 발현물을 C형 간염환자의 혈청으로 웨스턴 블롯팅한 결과를 나타낸 것이며,A of FIG. 3 shows the results of confirming the expression of the non-structural 5-1.2 gene by SDS polyacrylamide gel electrophoresis, and B of FIG. 3 shows the expression of the non-structural 5-1.2 gene as the serum of hepatitis C patients. Western blotting results are shown.

제 4 도의 A는 403 단백질 및 비구조 5-1.2 단백질의 발현을 SDS 폴리아크릴아미드겔 전기영동으로 확인한 결과를 나타내며4A shows the results of confirming the expression of 403 protein and non-structural 5-1.2 protein by SDS polyacrylamide gel electrophoresis.

제 4 도의 B는 403 및 비구조 5-1.2 유전자의 발현물을 C형 간염 환자의 혈청으로 웨스턴 블로팅하여 비교한 결과를 나타낸다FIG. 4B shows the result of Western blotting of the expressions of the 403 and nonstructural 5-1.2 genes with serum of hepatitis C patients.

Claims (8)

유비퀴틴에 하기 아미노산 서열을 갖는 한국형 C형 간염 바이러스의 비구조 5-1.2 단백질이 융합된 재조합 단백질.A recombinant protein in which ubiquitin is fused with a non-structural 5-1.2 protein of a Korean hepatitis C virus having the following amino acid sequence. 제 1 항의 재조합 단백질을 코딩하는 뉴클레오티드 서열을 갖는 DNA.DNA having a nucleotide sequence encoding the recombinant protein of claim 1. 제 2 항에 있어서, 한국형 C형 간염 바이러스의 비구조 5-1.2 단백질을 코딩하는 하기 뉴클레오티드 서열을 포함하는 DNA.The DNA of claim 2 comprising the following nucleotide sequence encoding the nonstructural 5-1.2 protein of the Korean hepatitis C virus. 제 2 항 또는 제 3 항의 DNA를 포함하는 발현벡터.An expression vector comprising the DNA of claim 2 or 3. 제 4 항에 있어서, ptrpH-UB-NS5-1.2인 발현벡터.The expression vector according to claim 4, which is ptrpH-UB-NS5-1.2. 제 4 항 또는 제 5 항의 발현벡터로 형질전환된 대장균.E. coli transformed with the expression vector of claim 4 or 5. 제 6 항에 있어서, ptrpH-UB-NS5-1.2로 형질전환된 대장균.The Escherichia coli transformed with ptrpH-UB-NS5-1.2. 제 6 항 또는 제 7 항의 대장균을 배양함을 포함하는, 유비퀴틴에 융합된 C형 간염 바이러스의 비구조 5-1.2 유전자 단백질의 제조 방법.A method for producing a non-structural 5-1.2 gene protein of hepatitis C virus fused to ubiquitin, comprising culturing E. coli of claim 6 or 7. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019930026611A 1993-03-18 1993-12-06 Process for producing nonstructural 5-1.2 protein of hepatitis c virus(hcv) KR100285585B1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
KR1019930004165 1993-03-18
KR1019930004165A KR940021733A (en) 1993-03-18 1993-03-18 Method for preparing non-structural 5-1.2 protein of hepatitis C virus
KR93-4165 1993-03-18

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KR940021575A true KR940021575A (en) 1994-10-19
KR100285585B1 KR100285585B1 (en) 2001-04-02

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KR1019930026611A KR100285585B1 (en) 1993-03-18 1993-12-06 Process for producing nonstructural 5-1.2 protein of hepatitis c virus(hcv)

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