摘要
参考GenBank上已发表的胞内劳森氏菌相关蛋白序列,设计了4对引物,分别扩增了4个抗原性候选基因(3个外膜蛋白基因和1个表面脂蛋白基因),并将4个抗原性候选基因构建好原核表达载体后进行原核表达、SDS-PAGE电泳和Western blotting分析。SDS-PAGE电泳检测初步确定重组融合蛋白pET32a-LI0902、pET32a-LI1022能够进行表达,分别得到53、37kDa的条带;Western blotting分析则初步确定pET32a-LI1022具有抗原性。研究结果首次验证胞内劳森氏菌外膜蛋白基因是否具有抗原性,为诊断试剂盒及基因工程疫苗的研制提供理论基础。
According to the associated protein sequences of Lawsonia intracellularis published in GenBank, four pairs of primers were designed and the genes of three outer membrane proteins and one eetal lipoprotein were amplified. After constructing the prokaryotic expression vectors for four antigenic candidate genes, the prokaryotic expression, SDSPAGE eleetrophoresis and Western blotting analysis were performed. The results showed that two recombination fusion proteins pET32a-LI0902 and pET32a-LI1022 could be expressed and 57kDa and 37kDa band was obtained by SDS-PAGE electrophoresis, respectively. The Western blotting analysis results showed that pET32a-LI1022 had antigenieity. These resuits for the first time proved that whether outer membrane proteins of Lawsonia intraeellulafs had antigenieity or not, and would provide theoretical basis for developing diagnostic kit and genetically engineered vaccine.
出处
《广西农业科学》
CAS
CSCD
2010年第1期62-65,共4页
Guangxi Agricultural Sciences
基金
广西科学研究与技术开发计划项目(桂科能0630006-6B)
