Optimization of prokaryotic expression condition and purification of soluble GST-CRH protein
10.11958/20161302
- VernacularTitle:可溶性GST-CRH蛋白原核表达条件的优化及纯化
- Author:
Shuo YU
;
Feng CHEN
;
Yingfu LIU
;
Jingrui HUO
;
Guangzong LI
;
Yi ZHANG
;
Hui DING
;
Haojun FAN
- Keywords:
corticotropin-releasing hormone;
glutathione s-transferase;
prokaryotic expression;
purification
- From:
Tianjin Medical Journal
2017;45(2):146-150
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain the recombinant corticotropin releasing hormone (CRH) protein with soluble, high purity protein through optimizing prokaryotic expression condition and purifying glutathione thiol transferase (GST)-CRH protein. Methods To detect the expression of soluble CRH protein through grope of the host strain GST-CRH temperature of induction expression, the host strain concentration (OD600), IPTG concentration and induction time, the purification of GST-CRH was performed by GST-CRH agarose gel. Western Blot assay was used for the expression identification of the target protein. Results The optimal conditions for the induction of CRH protein were determined: temperature of 30 ℃, IPTG induced concentration 0.1 mmol/L, bacteria density (OD600) 0.8, the induction time of 8 hours, purified GST-CRH>95% fusion protein was obtained. Conclusion The optimal expression conditions of GST-CRH are obtained, and the soluble protein of high purity GST-CRH is also obtained.
