幽门螺杆菌vacA基因重组表达的包涵体复性及ELISA方法的建立
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党双锁, 王宏仓, 贾晓黎, 袁利超, 王宝峰,
幽门螺杆菌;vacA基因;包涵体;变性;复性,党双锁,王宏仓,贾晓黎,袁利超,王宝峰,张欣,张正国,程延安,通讯作者:,Renatu
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党双锁, 王宏仓, 贾晓黎, 袁利超, 王宝峰, 张欣, 张正国, 程延安, 西安交通大学第二医院感染科 陕西省西安市 710004
通讯作者: 党双锁, 710004, 陕西省西安市西五路157号, 西安交通大学第二医院感染科. shuangsuo640212@sohu.com
电话: 029-83036998
收稿日期: 2005-08-29 接受日期: 2005-09-06
Renaturation of Helicobacter pylori vacA gene recombined expressive inclusion body and construction of ELISA method
Shuang-Suo Dang, Hong-Cang Wang, Xiao-Li Jia, Li-Chao Yuan, Bao-Feng Wang, Xin Zhang, Zheng-Guo Zhang, Yan-An Cheng
Shuang-Suo Dang, Hong-Cang Wang, Xiao-Li Jia, Li-Chao Yuan, Bao-Feng Wang, Xin Zhang, Zheng-Guo Zhang, Yan-An Cheng, Department of Infectious Disease, the Second Hospital of Xi'an Jiaotong University, Xi'an 710004, Shaanxi Province, China
Correspondence to: Shuang-Suo Dang, Department of Infectious Disease, the Second Hospital of Xi'an Jiaotong University, 157 West 5th Road, Xi'an 710004, Shaanxi Province, China. shuangsuo640212@sohu.com
Received: 2005-08-29 Accepted: 2005-09-06
Abstract
AIM: To increase the biological activity of the expressive product through denaturation and renaturation of H pylori vacA gene recombined expressive inclusion body, and to discuss the feasibility of mass production of VacA antigen.
METHODS: The VacA antigen expressed engineering bacterium was induced by Isopropyl-β-D-thiogalactopy-
ranoside (IPTG). The expressed inclusion body was denatured, renatured and dialyzed, and its activity was identified. The plates of ELISA were folded by the produced antigen. Then the ELISA method was constructed to determine the biological activity of the antigen.
RESULTS: The recombined engineering bacterium expressed Mr 3 300 target protein in the form of in-clusion body. SDS-PAGE showed the purity of the optimized inclusion body was above 95%. The ac-cordant rate was 97.1% between ELISA and blot electrophoresis after the activity examination was performed in 126 healthy people ......
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