副猪嗜血杆菌外膜蛋白TbpB基因的克隆和表达  

Cloning and expression TbpB gene for outer membrane protein of Haemophilus parasuis

作　　者：叶飞[1,2] 张培君[1] 田德雨[3] 孙慧玲[1] 王宏俊[1] 龚玉梅[1] 贺云霞[1] 

机构地区：[1]北京市农林科学学院畜牧兽医研究所,北京100097 [2]首都师范大学生命科学学院,北京100048 [3]中国农业大学动物医学院,北京100094

出　　处：《黑龙江畜牧兽医》2011年第8期25-27,共3页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31001072);"863"国家高技术研究发展计划项目(2006AA10A206);北京市农林科学学院青年基金项目(QNJJ201012);北京市农林科学学院一般项目(2010A008)

摘　　要：为了克隆和表达副猪嗜血杆菌(Haemophilus parasuis,HPS)外膜蛋白TbpB基因,试验采用基因工程的方法对已发布的HPS外膜蛋白P5序列进行相关序列分析,设计并合成引物,以HPS血清5型基因组为模板,通过PCR扩增HPS外膜蛋白P5编码基因,获得长为1 116 bp的预期基因片段。该基因编码372个氨基酸的蛋白质,预测分子质量约为41 ku,将该基因片段定向克隆至表达载体pGE X-6p-1中。结果表明:诱导表达后分子质量约为67 ku,与副猪嗜血杆菌阳性血清发生了特异性反应。说明该蛋白与抗体发生了反应,有一定免疫原性。To study the immunogenieity of the proteins related with HPS, A pair of specific primers were designed and synthesized on the basis of the published genome as a template. The TbpB gene of HPS was amplified by PCR, and then was cloned into the expression vector pGEX - 6p - 1 . The fusion protein was successfully expressed, analyzed the immunogenicity of TbpB protein by the Western - blot. The results showed that the TbpB protein was obtained and its molecular weight was about 54 ku. The Western - blot results showed that it had immune response with the HPS positive serum and the protein had immunogenicity. It prepared a basis of subnnit vaccine and diagnostic reagents.

关 键 词：副猪嗜血杆菌 转铁结合蛋白B(TbpB) 克隆 表达 

分 类 号：S852.613[农业科学—基础兽医学]