中国生物工程杂志

Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software. Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template. The amplified fragments G1 and G2 are 570bp and 308bp in length, respectively. The PCR products were cloned into pET30a vector and expressed in soluble form in E. coli after induction of cultured E. coli with IPTG. Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions. Then the purified proteins were analysed by western blotting. The results showed that the purified recombinant protein G1 retained good antigenicity and specificity. But the purified recombinant protein G2 didn’t possess biological activity. Antibodies against BRSV were detected in suspected bovine sera by using indirect ELISA and western blotting with the purified recombinant protein G1 in China. The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection. And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.