遗传 ›› 1993, Vol. 15 ›› Issue (5): 6-10.

• 论文 • 上一篇    下一篇

蛋白质双向电泳、印迹转移及蛋白质合成无细胞体系技术在小鼠腹水细胞EF-3研究中的应用

王汝刚1;秦士良2 WANG Ru-Gang1;QIN Shi-Liang   

  1. 1.解放军总医院实验仪器中心,北京 100853 1.Instrument Centre of General Hospital PLA,Beijing 100853 2.北京师范大学生物第,100875 2.Department of Biology,Beijing Normal University,100875
  • 收稿日期:1900-01-01 出版日期:1993-10-10 发布日期:1993-10-10

The Application of the Methods of Two Dimentional Gel Electrophoresis,Western Blotting and Cell-free System of Protein Synthesis in the Research in EF-3 Factor of H22a Cell

  • Received:1900-01-01 Online:1993-10-10 Published:1993-10-10

摘要: 本文应用近年发展起来的生化技术――蛋白质双向电泳(其第一向为等电聚焦,第二向为SDS凝胶电泳),将小鼠腹水细胞核糖体蛋进行了指纹分离。并利用蛋白质印迹转移(Western blotting),将转移后的硝酸纤维膜与交联了碱性磷酸酶的第二抗体和抗酵母EF-3抗体反应,证实该核糖体蛋白含有EF-3同源片段,进而制备了蛋白质合成无细胞体系。通过测定PolyU指导下3 H-phe掺入活力的免疫失活实验,初步证实此同源片段是小鼠腹水细胞蛋白质合成所必需。
The ribosomal proteins of H22a cell,were separated with the method of two-D gel electrophoresis (the first dimention is isoelectric focus and the second is SDS PAGE).Then the Western blotting was used,the transferred nitrocellulose sheet was treated with antiyeast EF-3 antibody and the second antibody bonded with alklinephosphoesterase.The result shows that the ribosomal proteins have a homologous fragment to yeast EF-3 factor.The cell-free system of protein synthesis was also established.By determing the activity of polyU direeted 3H-phe intervention in immunodeactivitive experiment,it is primaril confirmed that this fragment is the esscntial for the protein biosynthesis in H22a cell.

关键词: 小鼠腹水细胞, EF-3因子, H22a cell, 同源片段
Key words,
无细胞体系