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Abstract

Expression on gB gene of simian B virus in insect cells.

Abstract

In order to obtain antigenic properties used to construct an ELISA kit to test a simian B virus (BV) antibody and take the place of the BV, the gB gene of the simian B virus/E2490 was synthesized according to the reported BV virus gB gene (GenBank accession number: AF533768). The gB gene fragment was cloned in the vector pFastBac-HTa. The recombinant plasmid pFastBac-HTa-gB was constructed and identified by PCR and enzyme digestion and was sequenced. pFastBac-HTa-gB was transformed into DH10Bac complement cells. It was identified by antibiotics and PCR, indicating that a Baculovirus expression vector to the gB gene was constructed. On this basis, transposition bacmid DNA was extracted to transfect sf9 insect cells. At 96 h after transfection, the sf9 cells expanded and Baculovirus was harvested. The SDS-PAGE, Western blot and indirect immunofluorescence assay results showed the expression of the gB protein in the insect cells. The gB protein had good biological activity and was approximately 98 kDa in length.