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人尿激酶原(pro-urokinase)基因在大肠杆菌中的高效表达
引用本文:马忠,华子春,董晨,夏炎,陈于红,朱德煦. 人尿激酶原(pro-urokinase)基因在大肠杆菌中的高效表达[J]. Acta biochimica et biophysica Sinica, 1995, 0(1)
作者姓名:马忠  华子春  董晨  夏炎  陈于红  朱德煦
作者单位:南京大学生物化学系,南京大学医药生物技术国家重点实验室
基金项目:国家“863”高技术发展计划资助
摘    要:本文用PCR方法对人工合成的尿激酶原。DNA基因5’端进行改造,将之克隆到表达载体pKK233-2中,转化大肠杆菌JA221,经IPTG诱导,获得了占总菌体蛋白15%的高表达。SDS-PAGE和Westernblot结果显示表达产物主要为单链尿激酶,并且在菌体内基本上以无活性的包含体形式存在。经体外交复性,从1升培养基中可获得300000单位的活性尿激酶原。

关 键 词:尿激酶原,纤溶作用,包含体

High Expression of Human Pro-urokinase Gene in Escherichia coli
MA Zhong,HUA Zi-Chun,DONG Chen,XIA Yan,OHEN Ye-Hong and ZHU De-Xu. High Expression of Human Pro-urokinase Gene in Escherichia coli[J]. Acta biochimica et biophysica Sinica, 1995, 0(1)
Authors:MA Zhong  HUA Zi-Chun  DONG Chen  XIA Yan  OHEN Ye-Hong  ZHU De-Xu
Abstract:Modified by PCR method at the 5' end of the gene, a chemically synthesized pro-urokinase(pro-UK) cDNA gene was eloned into the expression vector pKK 233-2,and was expressed in E.coli JA221 by IPTG induction. The expression level was 15% of total bacterial proteins.Results identified by SDS-PAGE and Western blot showed that the expressed product mainly consisted of the single chain pro-nrokinase,which existed in the form of inactive inclusion bodies.By denaturation find renaturation ac vitro,300000 units of active pro-urokinase were obtained from 1 L culture medium.
Keywords:Pro-urokinase  Fibrinolytic activity  Inclusion body  
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