Construction of Prokaryotic Plasmid of Various Kinase Activity Types of Human PAK6 Gene and Identification of Its Recombinant Protein

Objective To construct various kinase activity types of prokaryotic expression vector of PAK6(p21-activated kinase)gene and identify their recombinant protein expression induced by isopro-pyl-1-thio-b-Dgalactopyranoside(IPTG).Methods wild type,kinase active and kinase dead type pcDNA3.1-PAK6 were digested by EcoRⅠ and XhoⅠ,and they were cloned into pGEX-5X-1.The expressions of various types GST-hPAK6 fusion proteins were induced by IPTG and identified by Western blot.Results The wild type,kinase active and kinase dead types coding sequences of hPAK6 gene were cloned into the pGEX-5X-1 plasmid which was transformed into E.coli BL21,identified by double enzymes digestion and sequencing.The expression of GST-hPAK6 fusion protein was induced by IPTG,and the molecular weight of protein was 101 kDa.Conclusion The various typese of PAK6 recombinant prokaryotic plasmids were successfully constructed into pGEX-5X-1.The expression of GST-hPAK6 fusion proteins were induced by IPTG and identified.