结核分枝杆菌rBCG-Rv2029c重组疫苗的构建与鉴定  

Construction and identification of recombinant rBCG-Rv2029c vaccine of Mycobacterium tuberculosis

作　　者：薛士鹏[1] 吴建勇[1] 宋彬[1] 齐红双[2] 李英[2] 党颍徐 满永宏[1] 

机构地区：[1]南阳医学高等专科学校基础医学部,南阳473061 [2]河南大学附属南阳南石医院,南阳473006

出　　处：《中国人兽共患病学报》2017年第8期744-747,752,共5页Chinese Journal of Zoonoses

基　　金：校自科NYYZ006~~

摘　　要：目的构建结核分枝杆菌rBCG-Rv2029c重组疫苗并鉴定。方法通过PCR扩增Rv2029c抗原编码基因,然后用双酶切法将Rv2029c和pMV261质粒酶切,再将酶切产物连接成rpMV261-2029c重组质粒,用电穿孔法将该质粒导入BCG中构建成rBCG-Rv2029c重组疫苗,最后用SDS-PAGE和Western blotting鉴定表达的重组蛋白。结果通过PCR成功扩增出1 020bp的Rv2029c基因,插入到pMV261质粒中,再把融合基因成功导入BCG中,经双酶切及基因比对鉴定证实,再通过热诱导后用Western blotting显示重组蛋白具有免疫原性。结论成功构建了结核分枝杆菌rBCG-Rv2029c重组活疫苗,为重组疫苗的免疫机制研究奠定基础。We constructed a recombinant vaccine of Mycobacterium tuberculosis rBCG-Rv2029 c,and then identified it.Rv2029 cantigen encoding gene was amplified by PCR.The enzyme digestion products were ligated into rpMV261-2029 crecombinant plasmid,after double digestion of Rv2029 cand pmv261vector,and then we introduced the plasmid into BCG to construct rBCG-Rv2029 crecombinant vaccine by electroporation method.Finally,we analyzed the expression of the recombinant protein by SDS-PAGE and Western blotting.A total of 1 020 bp Rv2029cgene successfully amplified by PCR was inserted into the plasmid pmv261,then the fusion gene was successfully transduced into BCG.After identified by double enzyme digestion,confirmed by gene alignment and by thermally induced with Western blotting,the recombinant protein had a free primary.The recombinant live vaccine of M.tuberculosis rBCG-Rv2029 cis successfully constructed,which lay a foundation for the study of the immune mechanism of recombinant vaccine.

关 键 词：卡介苗 重组疫苗 结核杆菌 Rv2029c 

分 类 号：R378[医药卫生—病原生物学]