Prokaryotic expression and purification of porcine Sox6

Authors : Wanxue WEN, Xiaoling CHEN, Zhiqing HUANG, Meng XU, Daiwen CHEN, Bing YU, Jun HE, Junqiu LUO, Xiangbing MAO, Jie YU, Ping ZHENG, Hong CHEN

Pages : 915-921

View : 10 | Download : 7

Publication Date : 2016-12-01

Article Type : Research Paper

Abstract :Sox6, a member of the Sox insert ignore into journalissuearticles values(Sry-related HMG box); family of transcription factors, plays an important role in embryonic muscle development and adult fast myofiber maintenance. Here we report the expression and purification of a His-tagged version of recombinant porcine Sox6 insert ignore into journalissuearticles values(pSox6); in Escherichia coli. The recombinant pSox6 was expressed as a C-terminal fusion protein with His6 tag in the E. Coli Rosetta insert ignore into journalissuearticles values(DE3); host strain. The protein was purified by Ni affinity chromatography, yielding approximately 5 μg/mL. The protein was identified by western blot analysis and confirmed by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight insert ignore into journalissuearticles values(MALDI-TOF/TOF); mass spectrometer analysis. In vitro biological activity assay demonstrated that the refolded purified recombinant pSox6 downregulated MyHC I, Tnnt1, Tnnc1, and Tnni1 mRNA expressions in porcine myotubes, suggesting that it was active. The present study provides a reliable technique for the recombinant expression and purification of pSox6 protein.
Keywords : Porcine Sox6, expression and purification, Escherichia coli, identification, in vitro assay