The expression and purificiation of human Annexin Ⅴ overexpressed in E. coli and the detection of apoptosis
- VernacularTitle:人重组钙依赖性磷脂结合蛋白Ⅴ在大肠杆菌中的高效表达、纯化及细胞凋亡检测
- Author:
Chunhui DI
;
Qun SHI
;
Yingmei ZHANG
;
Cheng ZHAO
;
Yingcheng ZHONG
;
Dawu HUANG
;
Dalong MA
- Publication Type:Journal Article
- From:
Journal of Peking University(Health Sciences)
2001;33(2):167-169
- CountryChina
- Language:Chinese
-
Abstract:
Objective: in order to provide rapid and reliable method. Methods: Encoded Annexin Ⅴ cDNA was amplifyed from U937 cDNA libary by PCR and then subcloned into E coli expression vector. MS2-Annexin Ⅴ fusion protein could be overexpressed in E coli. The MS2 bacteria protein could be removed by thrombin digestion.The mature Annexin Ⅴ was obtained by ion exchange chromatography and the FITC labled Annexin Ⅴ could be used in the detection of apoptosis. Results:Up to 37% of the total bacterial proteins was rhAnnexin Ⅴ as showed by SDS-PAGE. The purification of Annexin Ⅴ is over 99%. The FITC labled Annexin Ⅴ could efficiently detect apoptosis. Conclusion: We successfully established the technique procedure of obtaining a large quantity of Annexin Ⅴ and provided the basic routine for popularizing the detection of apoptosis' with high effciency.
