2011•Zhongguo yufang shouyi xuebaoRequires access
Expression of glycoprotein of rabies virus in Bac-to-bac baculovirus expression system
Gong Miao-miao
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Abstract
The glycoprotein gene of rabies virus Flury LEP strain was amplified by RT-PCR and cloned into pFastBacHTa vector. The resultant recombinant plasmid was transformed into competent cells of E. coli DH10Bac to construct recombinant bacmid. The recombinant baculovirus was generate by transfection of recombinant bacmid into sf21 insect cells. Expression of the recombinant glycoprotein (rG) was detected by SDS-PAGE with a MW of 60 ku, which was recongized with both ant-his tag monoclonal antibody and rabies virus positive serum by western blot test. A indirect ELISA was establish with rG as coating antigen, comparing with a commercial ELISA kit coating rabies virus as antigen, tested on 36 clinic serum samples,the indirect ELISA had 80.6% coincidence.