Supported By The National Natural Science Foundation Of China

Objective To produce the recombinant core particle of the hepatitis B virus with baculovirus expression vector system.Methods HBV P gene and C gene were amplified by PCR and cloned into pFastBac Dual to construct the recombinant plasmid pFastbac Dual-P-C, using pMD18-T Simple Vector as a mediator. Then the recombinant plasmid pFastbac Dual-P-C was transformed into DH10Bac competent cells for transposition, after which the recombinant Bacmid DNA was isolated and transfected into Sf9 insect cells with the help of lipofectamine. Three days later, recombinant baculoviruses were harvested to infect Sf9 cells. Then the interest proteins were purified from the infected cells through co-immunoprecipitation. SDS-PAGE and RT-PCR were applied to detect the different ingredients in the purified product.Results The recombinant baculovirus containing HBV P gene and C gene was constructed and the core particle was expressed in insect cells successfully. The purified product contained HBV polymerase, core protein and RNA.Conclusion The HBV core particle was successfully expressed in insect cells with baculovirus expression vector system and easily purified, which lays the foundation for further research.