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    인간 알파 S1 카제인 유전자로 형질전환된 대장균
    1.
    发明授权
    인간 알파 S1 카제인 유전자로 형질전환된 대장균 失效
    人类ALPHA S1-CASEIN表达载体和重组人类ALPHA S1-CASEIN

    公开(公告)号：KR100266753B1

    公开(公告)日：2000-09-15

    申请号：KR1019980006237

    申请日：1998-02-26

    Applicant: 한국과학기술연구원

    Inventor： 유대열 ,  김유경 ,  정봉현 ,  노낼달보 ,  이경광

    IPC: C12N15/70 ,  C12N15/63

    Abstract: PURPOSE: Provided is E. coli transformant that is transformed with human alpha S1 casein gene, thereby the human alpha S1 casein can be effectively produced, so that it can be used in studies about the functions of the human alpha S1 casein. CONSTITUTION: The transformant E. coli BL21(DE3)/pHalpha S1C2(KCTC 8867P) is prepared by the steps of: amplifying cDNA of human alpha S1 casein by PCR using probes and treating it with endonucleases such as Ndel/SaII and Ndel/XhoI; inserting the amplified cDNA of human alpha S1 casein into plasmid pET 22b(+) to construct pHalpha S1C1 and pHalpha S1C2; and transforming E. coli BL21 with pHalpha S1C1 and pHalpha S1C2. The mino acid sequence of human alpha S1 casein is analyzed by the steps of: incubating E. coli BL21(DE3)/pHalpha S1C2(KCTC 8867P) and collecting the cells; crushing the cells, and followed by centrifuging and separating it into a water-soluble fragment and a water-insoluble fragment; subjecting the fragments to Ni-NTA column chromatography; and analyzing the amino acid sequence of the separated and purified human alpha S1 casein with a protein sequencer.

    Abstract translation: 目的：提供用人α1S酪蛋白基因转化的大肠杆菌转化体，从而可以有效地产生人α1S酪蛋白，从而可用于关于人α1S酪蛋白的功能的研究。 构成：通过以下步骤制备转化体大肠杆菌BL21（DE3）/ pHalpha S1C2（KCTC 8867P）：通过使用探针通过PCR扩增人α1S酪蛋白的cDNA并用核酸内切酶如Ndel / SaII和NdeI / XhoI ; 将人α1S酪蛋白的扩增cDNA插入质粒pET223（+）中以构建pHalpha S1C1和pHalpha S1C2; 并用pHalpha S1C1和pHalpha S1C2转化大肠杆菌BL21。 通过以下步骤分析人α1S酪蛋白的微型酸序列：温育大肠杆菌BL21（DE3）/ pHalpha S1C2（KCTC 8867P）并收集细胞; 粉碎细胞，然后离心分离成水溶性片段和水不溶性片段; 将片段进行Ni-NTA柱色谱; 并用蛋白测序仪分析分离和纯化的人α1S酪蛋白的氨基酸序列。

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