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《Journal of Henan Agricultural Sciences》 2009-12
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Expression and Purification of Hog Cholera Lapinised Virus E~(rns) Gene in Escherichia coli

WANG Li1,YANG Yan-yan1,ZHANG Hong2,LI Qing-zhou3,LI Xue-wu1,ZHANG Gai-ping1(1.Henan Provincial Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;2.Henan Educational Institute,Zhengzhou 450003,China;3.Agricultural Economicsand Information Research Center,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China)  
The full lengh of Erns gene was subcloned into the prokaryotic expression vector pGEX-6p-1.The results of PCR amplification,restriction enzyme digestion and DNA sequencing showed that the recombinant plasmid was obtained.Then it was transformed into E.coli BL21(DE3)competent cells for expression.The expressed protein was identified by SDS-PAGE and Western-Blot,which showed the molecular weight of Erns fusion protein was approximately 55KD.E.coli BL21(DE3) harboring the recombinant plasmid pGEX-6p-1-Erns expresses the protein at high level(45%).The expressed protein was specifically recognized by rabbit anti-CSFV polyclonal antibodies.Ernsprotein was purified by GST-chelating chromatography.This may provide tools for further development of diagnostic reagent for detecting CSF antibodies.
【Fund】: 国家“863”计划项目(2007AA100606)
【CateGory Index】: S852.5
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