Cloning and expression of VP7 gene of African horse sickness virus

Abstract

Abstract: To obtain the African horse sickness virus (AHSV) VP7 protein, which is used to preparing serological diagnostic reagent and constructing AHS new generation vaccine, the prokaryotic expression is used to express VP7 protein. The VP7 gene was synthesized from the database of GeneBank, then the VP7gene was cloned into pBAD/Thio-TOPO vector, and the recombinant plasmid was transferred into Escherichia coli TOP10. The recombinant plasmid was sequenced to confirm the correct sequences and the correct junctional orientations of the inserted VP7 gene. The results of SDS-PAGE revealed that the VP7 recombinant fusion protein was expressed in TOP10 in a high level and the optimal amount of the expressed fusion protein is being induced with L-arabinose at 0.2% concentration for 4 hours. It had a molecular mass of approximately 54.72 kDa. The results of Western blotting and indirect ELISA revealed that the fusion protein is able to react with ASHV VP7 monoclonal antibody and they suggest that the fusion protein has immunologically reactive activity.

CLC Number:

S852.65+2

Zeng Zhaowen, Hua Qunyi, Duan Gang, Zhou Xiaoli, Dong Jun, Yang Yunqing, Yin Shanglian，Xiang Xun, Chang Hua. Cloning and expression of VP7 gene of African horse sickness virus[J]. Chinese Agricultural Science Bulletin, 2006, 22(10): 49-49.

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