H5N1亚型禽流感病毒核衣壳蛋白(NP)基因的进化及原核表达  

Phylogenetic Analysis and Prokaryotic Expression of NP Gene from Subtype H5N1 of Avian Influenza Virus in Escherichia coli

作　　者：邬成业[1,2] 王爱萍[2] 郝慧芳[1] 史平玲[2] 游雷鸣[1] 李培培[2] 张改平[1] 

机构地区：[1]河南省农业科学院农业部动物免疫学重点开放实验室河南省动物免疫学重点实验室,河南郑州450002 [2]郑州大学生物工程系,河南郑州450001

出　　处：《华北农学报》2010年第3期19-22,共4页Acta Agriculturae Boreali-Sinica

基　　金：国家"973"计划专项(2005CB523200)

摘　　要：参考GenBank上H5N1亚型禽流感病毒(AIV)的核衣壳蛋白(NP)基因序列设计引物,PCR扩增NP基因,对其进行序列分析并构建分子进化树。将克隆的NP基因插入原核表达载体pET32a,在大肠杆菌中进行诱导表达,对诱导表达的NP重组蛋白进行纯化、Western-Blot鉴定及免疫原性分析。结果表明,克隆的NP基因与GenBank上发表的另外2个河南AIV毒株亲缘关系较近,诱导表达的NP重组蛋白主要以可溶性形式存在,分子量约为70 kDa,并且能够与H5亚型AIV阳性血清发生特异性反应,具有良好的抗原性。The Nucleocapsid Protein（NP） gene of Avian influenza virus（AIV） subtype H5 was amplified by PCR according to the published sequence,and its molecular evolution was analyzed.The coding sequence of NP was cloned into the prokaryotic vector of pET32a to express and obtain the recombinant protein in E.coli.The expressed NP was purified and identified with Western-Blot strategy.and its immunogenicity was further characterized with en-zyme linked immunosorbent assay.The result suggested that the cloned NP was evolutionary more similar to others of Henan.and the fusion protein was successfully expressed and mainly existed as soluble bodies,Also,the protein could be specifically recognized by the serum against H5 subtype of AIV.

关 键 词：禽流感病毒 H5亚型 核衣壳蛋白(NP) 原核表达 

分 类 号：Q78[生物学—分子生物学]