Expression of VP genes of avian adeno-associated virus in insect cells

In order to express three structural proteins of avian adeno-associated virus( AAAV) in an appropriate proportion in insect cells,one pair of specific primers were designed according to the published genome sequences of AAAV to amplify VP genes by PCR,and the amplified fragment was cloned into Baculovirus expression vector pFastBac1. Then the recombinant vector pFastBac-VP was transformed into DH10Bac Escherichia. coli,and the positive recombinant bacmid rBacmid-VP was selected after resistance and blue-white plaque screening. rBacmid-VP was transfected into the Sf9 insect cells by liposome,and the rBac-VP was acquired. SDS-PAGE analysis showed three structural proteins,VP1,VP2 and VP3,were expressed in Sf9 with the right molecular weights and the ratio of 1 ∶ 1 ∶ 10. The results of Western blot showed that the recombinant proteins could be recognized by antisera of VP3.