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茨城病毒VP7蛋白基因的克隆表达及
间接ELISA方法的建立
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张文龙1，2，刘光亮1，王君伟2，吴东来1
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(1.中国农业科学院 哈尔滨兽医研究所 兽医生物技术国家重点实验室，黑龙江 哈尔滨150001；
2.东北农业大学 动物医学学院，黑龙江 哈尔滨150030)
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摘要：
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采用RT-PCR方法扩增出了茨城病毒(IBAV)No.2株编码VP7蛋白的S7全长基因，并克隆入原核表达载体pET-28a(+)中，在E.coli BL21中表达。经Western-blot检测，表达的重组VP7蛋白具有良好的反应原性。以纯化的重组蛋白作为包被抗原，初步建立了检测IBAV血清抗体的间接ELISA方法。 |
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关键词：
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茨城病毒；S7基因；表达；纯化；间接ELISA |
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中图分类号：S 852.659.4：R 446.61
文献标识码：
A 文章编号：1673-4696(2007)11-0960-04 |
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Establishment of an indirect ELISA assay for detecting Ibaraki virus |
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ZHANG Wen-long1，2,LIU Guang-liang1,WANG
Jun-wei2,WU Dong-lai1 |
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(1.National Key Laboratory of Veterinary Biotechnology/Harbin Veterinary Research Institute,Chinese Academy of
Agricultural Sciences,Harbin 150001,China;2.College of Veterinary Medicine,Northeast Agricultural
University,Harbin 150030,China) |
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Abstract: An indirect ELISA assay for detecting Ibaraki virus was developed. The method used VP7 protein of Ibaraki Ⅱ virus (IBAV Ⅱ) as an identification marker. The VP7 protein was a surface protein responsible for the antigenicity of IBAV Ⅱ. S7 Gene of IBAV Ⅱ encoding the VP7 protein was cloned into the pET-28a(+) prokaryotic expression vector. The vector was expressed in Escherichia coli BL21 as a fusion protein. The recombinant VP7 protein was purified and used in the indirect ELISA for detecting serum antibodies against IBAV Ⅱ. The recombinant protein had excellent antigenicity. |
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Key words: Ibaraki virus;S7 gene;expression;purification；indirect ELISA |
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