Cloning and Prokaryotic Expression of p53 Gene

p53 cDNA was obtained from human peripheral blood lymphocytes by RT-PCR,and then the p53 fragments were inserted into pQE40 vector to construct the recombinant plasmid pQE40-p53.Furthermore,pQE40-p53 plasmids were transformed into E.coli M15,and then induced by 1 mmol/L IPTG to express p53 protein in E.coli M15 expression system.The recombinant p53 protein fused with 6His-tag was purified by Ni-NAT resin affinity chromatography,and then urea was removed with gradual dialysis.Western Blot confirmed that purified 6His-p53 recombinant protein was obtained.