棉铃虫病毒HaSNPVfp25k基因的克隆表达及抗体制备  

Molecular Cloning and Expression of the fp25k Gene of HaSNPV in E. coli and Preparation of Its Antiserum

作　　者：吴东[1] 邓菲[1] 胡志红[1] 袁丽[1] 孙修炼[1] 

机构地区：[1]中国科学院武汉病毒研究所分子病毒学实验室

出　　处：《中国病毒学》2004年第4期380-384,共5页Virologica Sinica

基　　金：国家自然科学基金项目(30270058;30025003);973项目(2003CB114202)

摘　　要：根据棉铃虫单核衣壳核多角体病毒(Helicoverpaarmigerasinglenucleocapsidnucleopolyhedrovirus,HaSNPV)fp25k基因的序列,设计引物,引入适当的酶切位点,利用PCR扩增出基因片段。将该基因片段克隆至原核表达载体pProEXHTb,经IPTG诱导,在大肠杆菌DH5α中获得了高效表达,表达产物的大小为32kDa。纯化蛋白产物免疫家兔制备抗血清。该抗血清可与原核表达的GST-FP25K融合蛋白及在感染的昆虫细胞中表达的FP25K蛋白发生特异性免疫反应。该抗体的获得为深入研究FP25K蛋白的功能提供了基础。The PCR product of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus HaSNPV fp25k gene was cloned into a prokaryotic expression vector pProEXHTb. After inducing with IPTG, the fp25k gene was successfully expressed in E.coli DH5α. The expression product was 32 kDa. This protein was purified and injected into rabbit to raise polyclonal antiserum. Western blot analysis using this antiserum showed the specific reaction to FP25K protein expressed in both prokaryotic with GST fusion and viral infected insect cells. The antibody made it possible to analysis the function this protein in infected insect cells.

关 键 词：棉铃虫单核衣壳核多角体病毒 基因 克隆 表达 抗体 杆状病毒 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]