KR970074931A - Adenylate kinase 2A gene and its nucleotide sequence isolated from human fetal hepatocytes and its expression vector - Google Patents

Adenylate kinase 2A gene and its nucleotide sequence isolated from human fetal hepatocytes and its expression vector Download PDF

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KR970074931A
KR970074931A KR1019960017039A KR19960017039A KR970074931A KR 970074931 A KR970074931 A KR 970074931A KR 1019960017039 A KR1019960017039 A KR 1019960017039A KR 19960017039 A KR19960017039 A KR 19960017039A KR 970074931 A KR970074931 A KR 970074931A
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gene
adenylate kinase
nucleotide sequence
expression vector
derived
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KR1019960017039A
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KR100220089B1 (en
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최인성
이영희
김재화
이인애
강호범
정태화
최용경
임종순
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박원훈
한국과학기술연구원
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1229—Phosphotransferases with a phosphate group as acceptor (2.7.4)
    • 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
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/04—Phosphotransferases with a phosphate group as acceptor (2.7.4)
    • C12Y207/04003—Adenylate kinase (2.7.4.3)

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Abstract

본 발명은 에너지 대사 과정에 중요한 역할을 하고 그의 유전적 결함에 의해 비구상성 용혈성 빈혈 등이 유발되는, 사람의 아데닐레이트 키나제 유전자, 그의 염기 서열 및 본 유전자를 포함하여 아데닐레이트 키나제를 대장균에서 대량 생산할 수 있는 발현 벡터에 관한 것이다.The present invention relates to an adenylate kinase gene, a nucleotide sequence thereof, and an adenylate kinase of the present invention, which contains an adenylate kinase gene, a nucleotide sequence thereof, and a gene of the present invention, which plays an important role in the energy metabolism process, To an expression vector capable of mass production.

Description

사람의 태아 간세포에서 분리한 아데닐레이트 키나제 2A 유전자와 그의 염기 서열 및 이의 발현 벡터Adenylate kinase 2A gene and its nucleotide sequence isolated from human fetal hepatocytes and its expression vector

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음Since this is a trivial issue, I did not include the contents of the text.

제1도는 사람의 아데닐레이트 키나제 2A (AK2A) 유전자를 포함한 cDNA 클론 HAK2A의 전체 염기 서열 및 이 염기 서열로부터 예상되는 효소 단백질이 아미노산 서열을 나타낸 것이다. 제2도는 소의 AK2A 유전자(BOVAK2A) 및 쥐의 AK2A 유전자(RATAK2)에서 예상되는 효소 단백질의 아미노산 서열과 사람의 AK2A(HAK2A) 유전자에서 예상되는 아미노산 서열간의 유사성을 비교하여 나타낸 것이다. 제3도는 사람의 태아간 조직에서의 아데닐레이트 키나제의 발현을 노던블럿으로 분석한 것이다. 제4도는 대장균에서 사람의 AK2A 유전자를 발현하여 아데닐레이트 키나제를 생산할 수 있는 발현 벡터의 제조 과정을 나타낸 것이다. 제5도는 대장균에서 발현된 사람의 AK2A 유전자 산물이 아데닐레이트 키나제의 활성을 가짐을 나타낸 것이다.Fig. 1 shows the entire nucleotide sequence of the cDNA clone HAK2A containing the human adenylate kinase 2A (AK2A) gene and the amino acid sequence of the enzyme protein predicted from this nucleotide sequence. FIG. 2 shows the similarity between the amino acid sequence of the enzyme protein expected in the bovine AK2A gene (BOVAK2A) and the rat AK2A gene (RATAK2) and the amino acid sequence expected in human AK2A (HAK2A) gene. FIG. 3 is a Northern blot analysis of the expression of adenylate kinase in human fetal liver tissue. FIG. 4 shows a process for producing an expression vector capable of producing adenylate kinase by expressing human AK2A gene in Escherichia coli. FIG. 5 shows that the AK2A gene product of a human expressed in Escherichia coli has an activity of adenylate kinase.

Claims (10)

인체 조직 유래의 아데닐레이트 키나제 2A의 유전자.Genes of human tissue-derived adenylate kinase 2A. 제1항에 있어서, 아데닐레이트 키나제 2A 전체 유전자를 포함하는 cDNA 클론 HAK2A에서 유래한 것을 특징으로 하는 아데닐레이트 키나제 유전자.The adenylate kinase gene according to claim 1, wherein the adenylate kinase gene is derived from a cDNA clone HAK2A comprising the entire gene of adenylate kinase 2A. 제2항의 cDNA 클론 HAK2A로 대장균을 형질전환하여 제조하는 것을 특징으로 하는 형질전환체(수탁번호 : KCTC 8735P).A transformant (accession number: KCTC 8735P), which is produced by transforming E. coli with the cDNA clone HAK2A of claim 2. 서열번호 5의 염기 서열을 갖는 인체 조직 유래의 아데닐레이트 키나제 2A 유전자의 염기 서열.A base sequence of adenylate kinase 2A gene derived from human tissue having the nucleotide sequence of SEQ ID NO: 5. 제3항에 있어서, 제3항의 염기 서열의 전체 또는 10개 이상의 염기로 구성되고 AK2A 유전자와 70% 이상의 상동성을 가지고 있어서 불특정 다수의 유전자들 중에서 AK2A 유전자를 특이적으로 구분해 낼 수 있는 염기 서열을 가지고 있는 것을 특징으로 하는 제3항의 염기 서열 중 일부로 구성된 인체 조직 유래의 아데닐레이트 키나제 2A의 유전자의 염기 서열.[Claim 4] The method according to claim 3, wherein the nucleotide sequence of the nucleotide sequence of claim 3 or a nucleotide sequence which is composed of at least 10 bases and has 70% or more homology with the AK2A gene so as to specifically discriminate the AK2A gene from an unspecified number of genes A nucleotide sequence of a gene of adenylate kinase 2A derived from human tissue comprising a part of the nucleotide sequence of claim 3. 서열번호 6의 아미노산 서열을 갖는 인체 조직 유래의 아데닐레이트 키나제 2A 유전자의 아미노산 서열.An amino acid sequence of adenylate kinase 2A gene derived from human tissue having the amino acid sequence of SEQ ID NO: 6. 사람의 아데닐레이트 키나제 2A 유전자 또는 그 유전자의 일부를 포함하는 것을 특징으로 하는 발현 벡터.An expression vector characterized by comprising human adenylate kinase 2A gene or a part of the gene. 제6항에 있어서, 발현 벡터는 pHT-AK2A인 것을 특징으로 하는 발현 벡터.7. The expression vector according to claim 6, wherein the expression vector is pHT-AK2A. 제7항의 발현 벡터로 대장균을 형질전환하여 제조한 것을 특징으로 하는 형질전환제.A transformant produced by transforming E. coli with the expression vector of claim 7. 제9항의 형질전환체를 배양하여 아데닐레이트 키나제를 대장균에서 대량 생산하는 것을 특징으로 하는 효소 단백질의 제조 방법.The method for producing an enzyme protein according to claim 9, wherein the transformant is cultured to mass-produce an adenylate kinase in E. coli. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: It is disclosed by the contents of the first application.
KR1019960017039A 1996-05-20 1996-05-20 Gene for adenylate kinase 2a isolated from human fetal liver cells, nucleic acid sequence and expression vector thereof KR100220089B1 (en)

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