幽门螺杆菌HspA亚基的表达与纯化

Expression and purification of recombinant heat shock protein A subunit of Helicobacter pylori

AIM: To express and purify recombinant heat shock protein A subunit (HspA) of Helicobacter pylori (Hpyloi). METHODS: Gene hspA was amplified by PCR, and inserted into the prokaryotic expression vector pET22b, pQE-60 and pGEX-4T-2 respectively. The plamids were transformed into the Escherichia coli JM109 and BL21 (DE3). Expression of hspA gene was induced by IPTG and was analyzed by SDS-PAGE. The recombinant proteins were purified through nickel-affinity chromatography. Antigenicity of the recombinant proteins was analyzed by western blot. RESULTS: The hspA gene was expressed in two forms (HspA and HspA-His6) in pQE-60. The expressed protein accounted for above 40% of the total bacterial protein. The purity of HspA and HspA-His6 were 88.63% and 86.32% respectively after purification. Western blot proved that the recombinant proteins could be recognized by the anti-H pylori serum. CONCLUSION: HspA of H pylori has been expressed and purified with high efficiency, which can be used for vaccine development and immunological investigation.