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犬瘟热病毒水貂株H与F蛋白基因原核表达重组质粒的构建及表达

DOI: 10.7671/j.issn.1001-411X.2012.03.029, PP. 417-420

Keywords: 犬瘟热病毒,H基因,F基因,原核表达

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Abstract:

将克隆质粒pMD-18T-H和pMD-18T-F分别进行双酶切,获得纯化的H基因和F基因,在T4DNA连接酶的作用下,亚克隆到原核表达载体pET28a(+)上,获得原核重组质粒pET28a-H和pET28a-F.将原核重组质粒转化大肠埃希菌EscherichiacoliRosetta2(DE3)中进行表达,SDS-PAGE结果显示H基因和F基因分别表达相对分子质量约为31400和38200的融合蛋白,Western-blot检测结果显示,表达蛋白均可与CDV标准阳性血清呈阳性反应,表明原核表达的CDVH蛋白和F蛋白在反应原性上具有与天然H蛋白和F蛋白同样的特性,可作为CDV诊断用抗原,为CDV的免疫预防研究奠定基础.

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