副猪嗜血杆菌外膜蛋白pilA基因的克隆与表达  

Cloning and Expression of pilA Gene of 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,北京100193

出　　处：《安徽农业科学》2011年第13期7837-7838,7867,共3页Journal of Anhui Agricultural Sciences

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

摘　　要：[目的]克隆和表达副猪嗜血杆菌菌毛蛋白pilA基因。[方法]对已发表的HPS的pilA序列进行分析,合成引物,并以HPS血清5型基因组为模板,通过PCR扩增HPS的pilA编码基因,获得目的基因片段;构建重组表达质粒,经IPTG诱导表达至大肠杆菌BI21(DE3)中,进行SDS-PAGE与Western blot检测。[结果]表达的重组蛋白分子质量与预期的43 kD一致。[结论]该研究为制备亚单位疫苗和诊断试剂奠定了基础。[Objective] The aim was to clone and express the pilA gene of Haemophilus parasuis.[Method] The published pilA gene sequence of HPS was analyzed for primer synthesis,and the genome of serotype 5-type of HPS was used as template for PCR amplification of the pilA gene of HPS;the recombinant expression plasmid was constructed and transformed into E.coli BL21（DE3） after induced by IPTG.SDS-PAGE and Western blot analysis were then carried out.[Result] The molecular weight of expressed protein was consistent with the expected（43 kD）.[Conclusion] The results provided a foundation for the preparation of subunit vaccine and diagnostic reagents.

关 键 词：副猪嗜血杆菌 pilA 克隆 表达 

分 类 号：S188[农业科学—农业基础科学]