表达结核杆菌ppe27基因重组耻垢杆菌的构建及鉴定  

Construction and identification of recombinant Mycobacterium smegmatis expressing ppe27 gene of Mycobacterium tuberculosis

作　　者：于梦梦 张鑫逸 朱春霞 孙林 焦新安 

机构地区：[1]扬州大学江苏省人兽共患病学重点实验室/江苏省重要动物疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2015年第4期1-4,21,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家重点基础研究发展计划项目(973-2012CB518805);国家自然科学基金资助项目(31201882);江苏省高校自然科学基金资助项目(12KJB230001)

摘　　要：采用PCR技术扩增出结核分枝杆菌ppe27基因,并克隆至穿梭质粒pMV261中,将重组pMV261-ppe27-flag穿梭表达载体电转化到耻垢分枝杆菌内,诱导重组耻垢分枝杆菌,通过免疫印迹分析表达产物。结果表明:成功构建表达结核分枝杆菌基因ppe27的重组耻垢分枝杆菌,Western blotting结果显示,表达的重组融合蛋白PPE27-FLAG能与抗FLAG多抗以及兔抗PPE27-HIS蛋白多抗发生特异反应,在37.9ku处有明显特异性条带。表达ppe27基因的重组耻垢分枝杆菌构建成功,为下一步研究结核分枝杆菌致病机制奠定基础。The ppe27 gene was amplified from the genomic DNA of Mycobacterium tuberculosis strain H37 Rv and cloned into the shuttle plasmid pMV261.The recombinant plasmid pMV261-ppe27-flag was transformed into Mycobacterium smegmatis strain mc2 155 by electroporation.After heating inducing,PPE27-FLAG protein was expressed.Western blotting assay indicated that the recombinant protein could specifically react with anti-FLAG antibodies and polyclonal antibodies against PPE27-HIS protein respectively in 37.9ku.In conclusion,the recombinant M.smegmatis was successfully constructed and pave the way for further study of the pathogenesis of Mycobacterium tuberculosis.

关 键 词：结核分枝杆菌 ppe27基因 耻垢分枝杆菌 表达 

分 类 号：R378.91[医药卫生—病原生物学]