Construction and Prokaryotic Expression of Fusion Gene E6 and Mycobacterium Tuberculosis HSP70

Objective To construct fusion gene E6 and mycobacterium tuberculosis HSP70.The fusion gene was expressed in E.coli.and the fusion protein was purified.Methods The E6 and HSP70 gene were amplified by PCR and were cloned into expression vector pGEX-4T-1 after being sequenced.The DH5α contained this expression vector was induced by IPTG.The fusion protein was purified with GSTrap FF column.Results The E6 and HSP70 gene were amplified successfully and their sequence were identical to the reported in the GeneBank.The fusion gene PGEX-E6-HSP70 was constructed successfully.A GST fusion protein of 113 kD was detected with anti GST antibody by Western blotting.The SDS-PAGE showed that most of recombinant protein was soluble.Conclusion The fusion protein E6 and HSP70 was expressed and purified successfully.Which prepare the materials for further research of the fusion protein as a vaccine.