恒河猴凝血因子VII的体外表达初探  

STUDY OF THE IN VITRO EXPRESSION SYSTEM OF COAGULATION FACTOR VII OF RHESUS MONKEY,MACACA MULATTA

作　　者：王建东[1] 丁杨[1] 谭卫东[1] 陈又南[1] 陆燕蓉[1] 卢晓风[1] 李宏霞 王莉 程惊秋[1] 

机构地区：[1]四川大学华西医院卫生部移植工程与移植免疫重点实验室,成都610041 [2]成都(国家)中药安全评价中心

出　　处：《现代预防医学》2008年第13期2474-2475,共2页Modern Preventive Medicine

基　　金：高等学校博士学科点专项科研基金资助课题项目(20050610099)

摘　　要：[目的]探索恒河猴凝血因子VII的体外原核蛋白表达方法。[方法]利用已得到的恒河猴凝血因子VII cDNA序列,构建了带有6His表达标签的大肠杆菌原核表达重组质粒pET-DsbA-MCFVII,再转入大肠杆菌中进行诱导表达,并使用SDS-PAGE蛋白质电泳和Western Blot法检测蛋白表达情况。[结果]目的蛋白主要以包涵体形式表达,但是在菌液上清和菌体裂解液中也能检测到可溶形式的目的蛋白。[结论]恒河猴凝血因子VII在大肠杆菌中能够以可溶形式表达。[ Objective] To gain an insight into the in vitro prokaryofic expression profile of rhesus Factor Ⅶ in E. coll. [ MethOds] The recombinant prokaryotic E. coli expression vector pET-DsbA-MCFVII was successfully constructed, and then expressed in E. coll. The products were checked by SDS-PAGE and Western-Blot with anti-His tag antibody. [Results] Result showed Factor Ⅶ protein was mainly expressed as inclusion bodies, but little as soluble could be detected in supernatant. [Conclusion] Factor Ⅶ protein of rhesus monkey could be exoressed as soluble in E. coli.

关 键 词：恒河猴 凝血因子Ⅶ 原核表达 

分 类 号：R541.4[医药卫生—心血管疾病] R373.21[医药卫生—内科学]