Construction of Prokaryotic Plasmid of Human PAK4 Gene and Identification of Its Recombinant Protein

Objective To construct prokaryotic expression plasmid of human p21-activated kinase 4(hPAK4) gene and identify its recombinant protein expression induced by isopro-pyl-1-thio-b-Dgalactopyranoside(IPTG).Methods The coding sequence of PAK4 gene was digested from the plasmid(pCAN_(2)-PAK4_()) with XhoI and BamHI enzymes,and cloned into pGEX-5X-1.The expression of GST-hPAK4 fusion protein was induced by IPTG and identified by Western blot.Results The coding sequence of hPAK4 gene was cloned into the pGEX-5X-1 plasmid which was transformed into E.coli BL21.The length of the fragment was 2.4kb,identified by double enzymes digestion.The expression of GST-hPAK4 fusion protein was induced by IPTG,and the molecular weight of protein was 92KD.Conclusion The recombinant prokaryotic plasmid was successfully constructed into pGEX-5X-1.The expression of GST-hPAK4 fusion protein was induced by IPTG and identified.