Construction of GST-hPlk2 fusion protein expression vectors and the expression in prokaryotic cells

Objective To construct GST-hPlk2 fusion protein expression vector and induce its expression in Escherichia coli(E.coli).Methods Total mRNA was extracted from HEK293 cells,and cDNA was formed by reverse transcription.The hPlk2 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 prokaryotic expression plasmid pGEX-4T-2-hPlk2 was successfully constructed and confirmed by enzyme digestion and sequencing.The GST-hPlk2 fusion proteins were expressed and confirmed by Western blot.Conclusions The prokaryotic expression plasmid of hPlk2 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 Plk2 protein and the biological function of Plk2.