KR900001845A - Method for preparing hepatitis B virus surface antigen by yeast expression vector using complex promoter - Google Patents

Method for preparing hepatitis B virus surface antigen by yeast expression vector using complex promoter Download PDF

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KR900001845A
KR900001845A KR1019880008226A KR880008226A KR900001845A KR 900001845 A KR900001845 A KR 900001845A KR 1019880008226 A KR1019880008226 A KR 1019880008226A KR 880008226 A KR880008226 A KR 880008226A KR 900001845 A KR900001845 A KR 900001845A
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South Korea
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surface antigen
promoter
yeast
expression vector
hepatitis
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KR1019880008226A
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Korean (ko)
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KR900005534B1 (en
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박만훈
김성진
김경호
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허영섭
재단법인목암생명공학연구소
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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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity

Abstract

내용 없음.No content.

Description

복합 프로모터를 이용한 효모 발현 벡터에 의한 B형 간염 바이러스 표면항원들의 제조방법Method for preparing hepatitis B virus surface antigen by yeast expression vector using complex promoter

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

제1도는 pBR322 플라스미드의 BamHI부위에 HBV DNA가 클로닝된 pHBV 156 플라스미드를 나타내며,1 shows a pHBV 156 plasmid cloned with HBV DNA at the BamHI site of pBR322 plasmid,

제2도는 디데옥시 사슬 정지 DNA 서열방법에 의해 결정된 pre-S2부위 및 S유전자의 염기 배열 순서도를 나타내며,Figure 2 shows the sequence sequence of the pre-S2 site and the S gene determined by dideoxy chain stop DNA sequencing method,

제3도는 pre-S2부위와 S유전자의 5' 및 3' 말단의 불필요한 부분을 제거하여 코딩 서열만을 갖는 pHB-s148 및 pHBPs149의 제조방법을 설명한 도면.3 is a diagram illustrating a method for preparing pHB-s148 and pHBPs149 having only coding sequences by removing pre-S2 and unnecessary portions of 5 'and 3' ends of the S gene.

Claims (14)

효모의 α-인자의 조절부분과 갈락토키나제 조절부분을 융합시킨 프로모터를 이용함을 특징으로 하여 B형 간염 바이러스의 표면항원을 효모에서 형질발현 할 수 있는 서열을 함유한 재결합체 발현벡터.Recombinant expression vector containing a sequence capable of expressing the surface antigen of hepatitis B virus in yeast by using a promoter in which the regulatory portion of the α-factor of the yeast and the galactokinase regulatory portion are fused. 제1항에 있어서, B형 간염 바이러스 표면항원을 코오딩하는 서열이 adr형의 S유전자만을 포함하는 발현벡터.The expression vector of claim 1, wherein the sequence encoding the hepatitis B virus surface antigen comprises only an adr type S gene. 제1항에 있어서, B형 간염 바이러스 표면항원을 코오딩하는 서열이 adr형의 pre-S2 부위 및 S유전자를 포함하는 발현벡터.The expression vector of claim 1, wherein the sequence encoding the hepatitis B virus surface antigen comprises a pre-S2 region and an S gene of adr type. 효모의 α-인자의 조절부분과 칼락토키나제 조절부분을 융합시킨 프로모터를 이용함을 특징으로 하여 B형 간염 바이러스의 표면항원을 효모로부터 형질발현 할 수 있는 서열을 함유한 재결합체 발현 벡터의 제조방법.Method for producing a recombinant expression vector containing a sequence capable of expressing the surface antigen of hepatitis B virus from yeast, characterized by using a promoter in which the regulatory portion of the α-factor of yeast and the regulatory portion of the lactokinase are fused. . 제4항에 있어서, 표면항원 유전자들의 3'말단의 불필요한 부분을 제거하는 방법.The method of claim 4, wherein the unwanted portion of the 3 ′ end of the surface antigen genes is removed. 제4항에 있어서, 표면항원 유전자들의 5'말단의 불필요한 부분을 제거하는 방법.The method of claim 4, wherein the unwanted portion of the 5 ′ end of the surface antigen genes is removed. 제4항에 있어서, α-인자의 프로모터의 일부와 갈락토키나제 프로모터의 일부분을 Sau3A 부위를 이용하여 결합시키는 방법.The method of claim 4, wherein a portion of the α-factor promoter and a portion of the galactokinase promoter are joined using a Sau3A site. 제4항에 있어서, 혼성 프로모터와 표면항원 유전자를 재결합시키는 방법.The method of claim 4, wherein the hybrid promoter and the surface antigen gene are recombined. 제4항에 있어서, 표면항원 유전자의 3'말단에 효모 α-인자 정지서열을 연결하는 방법.The method of claim 4, wherein the yeast α-factor stop sequence is linked to the 3 ′ end of the surface antigen gene. 제4항에 있어서, 프로모터, 표면항원 유전자, 정지서열을 포함하는 DNA 절편을 효모 벡터를 옮기는 방법.The method of claim 4, wherein the DNA fragment comprising the promoter, the surface antigen gene, and the stop sequence is transferred to the yeast vector. 제4항 또는 7항에 있어서, α-인자의 프로모터를 Sau3A를 사용하여 분리하는 방법.8. The method of claim 4 or 7, wherein the promoter of the α-factor is separated using Sau3A. 제4항 또는 7항에 있어서, 갈락토키나제 프로모터 부위를 DdeI과 Sau3A를 사용하여 분리하는 방법.8. The method of claim 4 or 7, wherein the galactokinase promoter site is separated using DdeI and Sau3A. 효모의 α-인자의 조절 부분과 갈락토키나제 조절 부분을 융합시킨 프로모터를 이용하여 B형 간염 표면항원을 코딩하는 서열이 adr형의 S유전자만을 포함하는 발현 벡터로 형질 전환된 사카로마이세스.세레비지에 20B-12/PMH148 형질 전환체.Saccharomyces wherein the sequence encoding hepatitis B surface antigen is transformed with an expression vector containing only adr-type S gene using a promoter in which the regulatory portion of the α-factor of the yeast and the galactokinase regulatory portion are fused. 20B-12 / PMH148 transformants in cerevisiae. 효모의 α-인자의 조절 부분과 갈락토키나제 조절 부분을 융합시킨 프로모터를 이용하여 B형 간염 표면항원을 코딩하는 서열이 adr형의 pre-S2부위 및 S유전자를 포함하는 발현 벡터로 형질전환된 사카로마이세스.세레비지에 20B-12/PMH149 형질 전환체.Using a promoter in which the regulatory portion of the α-factor of the yeast and the galactokinase regulatory portion were fused, the sequence encoding the hepatitis B surface antigen was transformed into an expression vector containing the adr type pre-S2 region and the S gene. Saccharomyces. 20B-12 / PMH149 transformant in cerevisiae. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019880008226A 1988-07-02 1988-07-02 Producing method for hepatitis b virus surface antigen KR900005534B1 (en)

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