H5N1亚型禽流感病毒NA和NS1蛋白原核表达及多克隆抗体制备  

Prokaryotic expression of the NA and NS1 genes of H5N1 subtype aivian influenza virus and preparation of their polyclonal antibodies

作　　者：吴艳菊[1,2] 张治业[1,2] 倪小舒 孟菲菲[1,2] 秦涛[1,2] 陈素娟[1,2] 彭大新[1,2] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏省动物重要疫病与人兽共患防控协同创新中心,江苏扬州225009

出　　处：《黑龙江畜牧兽医》2017年第6期1-4,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31402229);江苏高校优势学科建设工程项目;江苏高校品牌专业建设工程资助项目;扬州市青年科技人才项目(YZ2014028);扬州大学科研创新项目(x2015722)

摘　　要：为了获得检测禽流感病毒神经氨酸酶(NA)蛋白和非结构蛋白(NS1)的抗体,试验采用扩增NA、NA+、NS1和NS1+基因,连接原核表达载体p ET-32a进行原核表达,蛋白纯化后免疫Balb/c小鼠,获得NA、NA+、NS1和NS1+的多克隆抗体血清。结果表明:NA、NA+、NS1和NS1+多克隆抗体血清反应性良好,可以检测NA和NS1自身蛋白,且NA和NA+及NS1和NS1+多克隆抗体血清之间均具有良好的交叉反应。说明基因部分缺失对整个NA蛋白和NS1蛋白的抗原性影响不大。In order to prepare the polyelonal antibodies of neuraminidase （ NA ） and non - structurall （ NS1 ） protein of HSN1 subtype AIV, the NA and NS1 genes were cloned into the prekaryotic vector pET -32a（ ＋ ）, respectively. Furthermore, the recombinant plasmid was trans- formed into E. coli BL21 cells for expression of the NA, NA ＋, NS1, or NS1 ＋ protein. The antisera against NA, NA ＋, NS1, or NS1 ＋ protein were generated in Balb/c mice by using purified protein as immunizing antigens. The results indicated that the prepared sera exhibited good reactivity when detected by the recombinant proteins of the NA , NA ＋, NS1 or NS1 ＋, respectively. Moreover, there was favorable cross - reactivity between NA antiserum and NA ＋ protein, or between NA ＋ antiserum and NA protein in Western - blot assay. Meanwhile, anti- sera against NS1 and NS1 ＋ also had the similar characteristics of cross - reaction.

关 键 词：H5N1亚型 禽流感病毒 NA基因 NS1基因 原核表达 多克隆抗体 

分 类 号：S852.659.5[农业科学—基础兽医学] Q816[农业科学—兽医学]