北京大学学报(自然科学版)

重组PAI-1在大肠杆菌中表达条件的研究及其对t-PA,u-PA抑制的分析

隋广超,张曼,胡美浩   

  1. 北京大学生命科学学院,北京,100871
  • 收稿日期:1996-02-03 出版日期:1996-11-20 发布日期:1996-11-20

Study of Expression Conditions of Recombinant PAI-1 in E. coli and the Analysis of PAI1 Inhibition Towards t-PA, u-PA

SUI Guangchao, ZHANG Man, HU Meihao   

  1. College of Life Sciences, Peking University, Beijing, 100871
  • Received:1996-02-03 Online:1996-11-20 Published:1996-11-20

摘要: 带有表达质粒pBV220/PAI-1的大肠杆菌中,在菌体密度为A600nm=0.3--0.4时诱导,表达重组PAI-1(rPAI-1)的效率最高。在诱导后的6h以内,rPAI-1的百分含量持续升高。rPAI-1经分离纯化并用盐酸胍激活后,能够与t-PA及u-PA反应,用SDS-PAGE和纤维蛋白自显影方法可以检测出它们所形成的复合物。

关键词: 重组PAI-1, 表达条件, 抑制活性

Abstract: Bacteria of Escherichia coli harboring the expression plasmid, pBV220/PAI-1, might express recombinant PAI-1(rPAI-1) at the highest effeciency when the bacteria were induced at the bacterial density of A600nm= 0.3--0.4. Furthermore, the percentage of rPAI-1 in the bacteria kept rising within the 6 hours after the induction. The expressed rPAI-1 activated by guanidine HCl could form complexes with t-PA or u-PA, which might be visualized by SDS-PAGE or fibrin autography.

Key words: recombinant PAI-1, expression condition, inhibitory activity

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