| 人尿激酶原（pro－urokinase）基因在大肠杆菌中的高效表达 |
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| 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].生物化学与生物物理学报,1995(1). |
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| 作者姓名： | 马忠 华子春 董晨 夏炎 陈于红 朱德煦 |
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| 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 |
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| 基金项目： | 国家“863”高技术发展计划资助 |
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| 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。
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| 关 键 词： | 尿激酶原，纤溶作用，包含体 |
High Expression of Human Pro-urokinase Gene in Escherichia coli |
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| 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(1). |
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| Authors: | MA Zhong HUA Zi-Chun DONG Chen XIA Yan OHEN Ye-Hong and ZHU De-Xu |
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| 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. |
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| Keywords: | Pro-urokinase Fibrinolytic activity Inclusion body |
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