高等学校化学学报

高等学校化学学报 ›› 1998, Vol. 19 ›› Issue (4): 564.

• 论文 • 上一篇    下一篇

丝心蛋白基因分子克隆与表达的初步探讨

李宏杰1, 蔡再生2, 施华芳1, 于同隐2, 黄伟达1   

  1. 1. 复旦大学生物化学系, 上海, 200433;
    2. 复旦大学高分子科学系, 上海, 200433
  • 收稿日期:1997-03-02 出版日期:1998-04-24 发布日期:1998-04-24
  • 通讯作者: 黄伟达
  • 作者简介:李宏杰,男,26岁,硕士.
  • 基金资助:

    国家自然科学基金

A Preliminary Study on Expression of Fibroin C Subunit as a Fusion Protein in E. Coli

LI Hong-Jie1, CAI Zai-Sheng2, SHI Hua-Fang1, YU Tong-Yin2, HUANG Wei-Da 1   

  1. 1. Department of Biochemistry, Fudan Univ., Shanghai, 200433;
    2. Department of Macromolecular Science, Fudan Univ., Shanghai, 200433
  • Received:1997-03-02 Online:1998-04-24 Published:1998-04-24

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被引次数

6

摘要： 通过聚合酶链反应(PCR)扩增丝心蛋白C亚基结构基因,并将其克隆到融合蛋白表达载体pRIT2T质粒中得到pRIT2T-FL质粒,在大肠杆菌株P2392内进行表达.十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹反应证明融合蛋白在大肠杆菌中得到了表达.

关键词: 丝心蛋白C亚基, 融合蛋白, ProteinA, 免疫印迹反应

Abstract: The structural gene of fibroin Csubunit(i.e. fibroin light chain, FL) was amplified by polymerase chain reaction(PCR) from pFL18 plasmid, which carries a cDNAclone of fibroin Csubunit. The PCR product was cloned on fusion protein expression vector pRIT2T, and was expressed in the E. Coli strain P2392. SDS-polyacrylamide gel electrophoresis(SDS-PAGE) and immunoblotting were carried out to verify the expression of protein Afibroin Csubunit fusion protein.

Key words: Fibroin C subunit, Fusion protein, Protein A, Immunoblotting

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