Abstract:In order to gain antigenic properties what was used to construct a ELISA kit to test BV antibody and to take the place of the BV. gB gene of Simian B Virus/E2490 was syntheticed according to the reported BV virus gB gene (genbank accession number: AF533768). The gB gene fragment was cloned in the vector pFastBacHTA. The recombinant plasmid pFastBacHTA- gB was constructed and was identified by PCR and enzyme digestion and sequenced. Then the plasmid pFastHTA- gB was transformed into DH10Bac complement cells. It was identified by antibiotics and PCR, it is shows we had constructed baculovirus expression vector to gB gene. On this basis, transposition bacmid DNA was extracted to transfect sf9 insect cells .After transfected 96 hours,the sf9 insect cells appear expansion、round and cell nucleus greaten ,and baculovirus was harvested .Then the SDS-PAGE and western bloting and indirect immunofluorescence assay showed that the expression of gB protein in insecet cells .The gB protein has a good biological activity and is approximate 98kDa . It is a base to construct a ELISA kit to test BV antibody. The result showed that we had constructed the recombinant plasmid pFastBacHTA- gB and baculovirus expression vector to gB gene , and the gB gene was expressed in insect cells .