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Prokaryotic expression and characteristics of nonstructural protein NS5 of goose Tembusu virus
Author(s): 
Pages: 1704-1709
Year: Issue:  10
Journal: Acta Agriculturae Zhejiangensis

Keyword:  goose Tembusu virusNS5 proteinprokaryotic expression;
Abstract: 通过在大肠杆菌中表达鹅坦布苏病毒非结构蛋白NS5对其纯化,并进行特性分析.根据鹅坦布苏病毒JS804株非结构蛋白NS5基因序列,设计1对特异性引物,利用PCR技术扩增得到全长NS5基因序列,并将其定向克隆至原核表达载体pET32a中,获得重组表达质粒pET32a-NS5,经双酶切鉴定及测序正确后,转化至大肠杆菌BL21(DE3)中.经IPTG诱导表达,表达的重组融合蛋白最终经SDS-PAGE及Western-blotting鉴定.结果显示,NS5融合蛋白分子质量约为120 ku,在诱导后5h达到表达量高峰,融合蛋白以包涵体形式存在,Western-blotting显示原核表达的蛋白可被兔抗鹅坦布苏病毒血清特异性识别,表明以NS5蛋白为抗原制备的多克隆抗体具有较高的敏感性与特异性.
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