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

Liu YuHong; Hua QunYi; Xu ZiZhong; Yang YunQing; Zhou XiaoLi; Dong Jun; Yin ShangLian; Xu JingYi; Gao Hong

Veterinary Science in China 36(9): 692-695

2006


Accession: 012847063

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Abstract
The H glycoprotein gene of the peste-des-petits-ruminants virus (PPRV), which was synthesized artificially according to the Indian vaccine, was subcloned from pUC18-H and inserted into the 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 a high level in Escherichia coli TOP10. The expressed fusion protein, which made up 10% of the total bacteria protein after being induced with 0.2 g/litre of L-arabinose for 5 h, was approximately 83 ku in molecular weight. In western blotting test, the protein could specifically react with the PPRV H monoclonal antibody, indicating that the prokaryotic expression of PPRV H gene could produce the H glycoprotein.