嗜肺军团菌mip/flaA二联优势抗原表位基因融合表达载体的构建及表达  

Construction and Expression of Vector with mip/flaA Advantages Epitope Genes of Legionella pneumophila

作　　者：何金蕾 彭冉[2] 张俊荣[1] 余泽英[1] 李娜丽莎 龚艳菊 陈莹[2] 陈达丽[1] 陈建平[1] 

机构地区：[1]四川大学华西基础医学与法医学院寄生虫学教研室,成都610041 [2]四川大学华西基础医学与法医学院,成都610041

出　　处：《四川大学学报（医学版）》2016年第1期1-6,共6页Journal of Sichuan University(Medical Sciences)

摘　　要：目的选取嗜肺军团菌mip和flaA优势抗原表位基因,构建mip/flaA二联优势抗原表位基因融合表达载体,并在原核系统中表达,为后续制备嗜肺军团菌蛋白疫苗提供初步的实验基础。方法运用生物信息学方法对Mip和FlaA蛋白的二级结构和表面特性如理化性质、亲水性、可塑性、抗原指数以及胞外区等方面进行分析,选择其活性表位可能存在的区域为优势抗原表位区。通过PCR扩增和T4连接酶构建pET-mip、pET-flaA和pET-mip/flaA优势抗原表位基因融合表达载体,并诱导其在大肠杆菌中表达。结果 Mip和FlaA都存在多个潜在的抗原表位位点,选取其优势抗原表位区域进行克隆和表达获得成功,并成功表达了mip/flaA二联优势抗原表位融合蛋白。结论 DNA Star软件和Expasy在线蛋白分析系统能够成功预测嗜肺军团菌Mip和FlaA抗原的表位;选取其优势抗原表位成功构建了pET-mip/flaA二联原核表达载体,并高效表达。Objective To generate and express fusion vector with mip/flaA advantages epitope genes of Legionella pneumophila by select mip and flaA advantages epitope genes for future research on Legionella pneumophila protein vaccine.Methods Following analysis of secondary structure and surface properties such as：physical and chemical properties,hydropathy,plasticity,antigen index and extracellular domain of Mip and FlaA proteins by bioinformatics methods,the region which active epitope may exist was selected as advantages epitope region.Then,the recombinant plasmid pET-mip,pET-flaA and pET-mip/flaA with advantages epitope genes were constructed by PCR amplification and T4 ligase connection,and induced the expression in E.coli.ResultsMany potential antigenic epitopes in Mip and FlaA were identified,and the selected advantages epitope regions were cloned and expressed successfully.Moreover,the mip/flaAtwo advantages associated epitope fusion proteins were also successfully expressed.Conclusion DNA Star software and Expasy online analysis system can successfully predict antigenic epitopes for Legionella pneumophila Mip and FlaA.And prokaryotic expression vector pET-mip/flaA with advantages epitope genes has been successfully constructed and efficiently expressed.

关 键 词：嗜肺军团菌 优势抗原表位 MIP基因 flaA基因 融合表达 

分 类 号：R378[医药卫生—病原生物学]