Tamdy病毒糖蛋白基因的重组表达及抗体制备Recombinant Expression of Glycoprotein Gene of Tamdy Virus and Antibody Preparation
刘利平;沈姝;刘希佳;张敏;王岚;何明宇;孙素荣;张渝疆;丁军涛;
摘要(Abstract):
原核表达Tamdy病毒(Tamdy virus,TAMV)糖蛋白Gn和Gc,分别制备兔抗融合蛋白Gn和Gc多克隆抗体。利用RT-PCR扩增Gn和Gc基因片段,分别构建原核表达质粒pET-28a-Gn和pET-32a-Gc,并在E.coli BL21(DE3)中诱导表达,镍柱亲和层析纯化融合蛋白Gn和Gc之后,以皮下注射方式分别免疫新西兰兔,制备兔抗融合蛋白Gn和Gc多克隆抗体。经Western Blotting和间接免疫荧光(IFA)鉴定多克隆抗体抗原识别能力,ELISA法测定抗体效价。结果显示,pET-28a-Gn、pET-32a-Gc原核表达质粒构建正确,融合蛋白Gn和Gc大小分别约为45 kD、74 kD,制备的兔抗融合蛋白Gn和Gc多克隆抗体能够分别识别原核表达的融合蛋白Gn和Gc和真核表达产物,效价分别为1∶409 600、1∶204 800。制备的多克隆抗体效价高,能够特异性识别原核系统和真核系统表达的TAMV糖蛋白,为TAMV的流行学分析提供基础。
关键词(KeyWords): Tamdy病毒;糖蛋白;原核表达;真核表达;蛋白纯化;多克隆抗体
基金项目(Foundation): 国家自然科学基金项目(项目号:81960369,81760365),题目:新型布尼亚样病毒(SFTSV-Like)结构蛋白(SP/GP)线性抗原表位及其自然疫源性研究,准噶尔盆地鼠类和媒介蜱携带重要虫媒病毒病原及其流行病学调查;; 新疆高校科研计划自然科学重点项目(项目号:XJEDU2019I002);题目:准噶尔盆地新疆出血热自然疫源地调查研究~~
作者(Authors): 刘利平;沈姝;刘希佳;张敏;王岚;何明宇;孙素荣;张渝疆;丁军涛;
DOI: 10.13242/j.cnki.bingduxuebao.004259
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