Tianjin Med J ›› 2017, Vol. 45 ›› Issue (2): 146-150.doi: 10.11958/20161302

• Cell and Molecular Biology • Previous Articles     Next Articles

Optimization of prokaryotic expression condition and purification of soluble GST-CRH protein

YU Shuo1, CHEN Feng2, LIU Ying-fu3, HUO Jing-rui2, LI Guang-zong2, ZHANG Yi2, DING Hui2, FAN Hao-jun1△   

  1. 1 Respiratory Department, the Affiliated Hospital of Xinxiang Medical University, Henan 453000, China; 2 Institute for Disaster and Emergency Rescue Medicine, the Affiliated Hospital of Logistics University of Chinese People’ s Armed Police Force; 3 Department of Cell Biology, Logistics University of Chinese People’ s Armed Police Force
  • Received:2016-11-08 Revised:2017-01-16 Published:2017-02-15 Online:2017-02-14
  • Contact: △Corresponding Author E-mail: fanhaojun999@126.com E-mail:978801515@qq.com

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.

Key words: corticotropin-releasing hormone, glutathione s-transferase, prokaryotic expression, purification