Construction of prokaryotic vector and expression of H glycoprotein gene of peste des petits ruminants virus

The H glycoprotein gene of peste des petits ruminants virus(PPRV),which was synthesized artificially according to the Indian vaccine,was sub-cloned from pUC18-H,and inserted into pBAD/Thio-TOPO vector.The recombinant plasmid was identified by PCR.Sequence analysis confirmed that the H glycoprotein gene was inserted correctly.SDS-PAGE analysis revealed that the H protein gene was expressed at high level in Escherichia coli TOP10.The expressed fusion protein,which made up 10% of total bacteria protein after being induced with 0.2g/L of L-arabinose for 5 hours,was approximately 83ku in molecular weight.In Western-blotting test,the protein can specifically react with the PPRV H monoclonal antibody,indicating that prokaryotic expression of PPRV H gene can produce H glycoprotein.