Expression,purification and transduction of PTD-p53 fusion protein to hepatocellular carcinoma cell

Objective To express and purify PTD-p53 fusion protein and investigate its transduction efficiency.Methods The gene encoding wide-type p53 was isolated,using RT-PCR from A549 cell line and cloned into pTATHA and pET32a prokaryotic expression vectors.Recombinant plasmids were E.coli BL21(DE3)LysS,then the transformed cells were induced with IPTG.The expression and purification of the PTD-p53 and p53 were analyzed by SDS-PAGE.BALB/c mice were immunized with purified p53 protein.The serum was isolated and the antibody specific to p53 was measured by ELISA.The transduction efficiency of PTD-p53 was detected using indirect immunoflure scence assay.Results Prokaryotic expression vectors of PTD-p53 and p53 were constructed correctly.PTD-p53 fusion protein and p53 protein were successfully expressed and purified.p53 specific mouse antiserum was obtained.IFA result indicated that PTD-p53 fusion protein transduced into HepG2 cells efficiently.Conclusion The obtained Tat-p53 fusion protein provides a theretical foundation for the basic research on PTD-p53 treating hepatocellular carcinoma.