To express mouse endostatin by thioredoxin fusion expression system
- VernacularTitle:用硫氧还蛋白融合表达系统表达小鼠内皮抑素的方法研究
- Author:
Weilie MA
;
Fang LIU
;
Chengwei HE
- Publication Type:Journal Article
- Keywords:
endostatin;
thioredoxin fusion expression system;
fusion protein
- From:Journal of Third Military Medical University
2003;0(14):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the best technology to produce recombinant mouse endostatin by thioredoxin fusion expression system.Methods The recombinant plasmid pThioHis-endo was further transformed into different E.coli.,including BL21,JM109,DH5?.After induction with IPTG of different concentration or for different time period,thioredoxin-endo fusion protein was expressed in E.coli.and the product was identified by SDS-PAGE.The recombinant endostatin expression and extraction,wash,degeneration and refolding of inclusion bodies were carried out by the optimized route,and the product was further purified by affinity chromatography through Ni~(+) column and identified by SDS-PAGE.Results No difference of endostatin expression in BL21,JM109,DH5? was found.The optimal concentration of IPTG was 0.9 mmol/L and optimal inductive phase was 5 h.The soluble recombinant endostatin could be obtained by affinity chromatography through Ni~(+) column.Conclusion Soluble endostatin recombinant fusion protein of high purity and yield could be obtained by the optimized technique route.
