High efficient expression of human endostatin in E.coli and its antiangiogensis activity
- VernacularTitle:人内皮抑素在大肠杆菌中的高效表达及活性研究
- Author:
Ge ZHANG
;
Xisong KE
;
Ailing DING
;
Wei YING
;
Kun YANG
;
Zheny ZHU
- Publication Type:Journal Article
- Keywords:
Endostatin;
Antibodies;
Biological activity
- From:
Chinese Journal of Pathophysiology
2000;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To examine the expression of human endostatin in E.coli , produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20 kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors.
