Construction of GST-hVinexin fusion protein expression vectors and the expression in prokaryotic cell

Objective To construct GST-hVinexin fusion protein expression vector and induce its expression in Escherichia coli(E.coli).Methods Total mRNA was extracted from COS-7 cells,and cDNA was formed by reverse transcription.The hVinexin coding sequence was amplified by polymerase chain reaction(PCR) and subcloned into pGEX-4T-2 vector.The positive recombinant was identified by restriction enzyme digestion and DNA sequencing.Then they were transformed into E.coli BL21,induced by IPTG and identified by SDS-PAGE and Western blot.Results The length of the fragment identified by restriction enzyme digestion was 990bp,the prokaryotic expression plasmid pGEX-4T-2-hVinexin was successfully constructed and confirmed by enzyme digestion and sequencing.The GST-hVinexin fusion proteins were induced and expressed positively by Western blot.Conclusion The prokaryotic expression plasmid of hVinexin was successfully constructed and the expression of fusion proteins in E.coli was confirmed.This study provides the basis for the further research on purifying Vinexin protein and the biological function of Vinexin.