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.
出处
《安徽农业科学》
CAS
北大核心
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)
关键词
副猪嗜血杆菌
pilA
克隆
表达
Haemophilus parasuis
pilA gene
Cloning
Expression