结核分枝杆菌rBCG-Rv2029c重组疫苗的构建与鉴定
发布时间：2019-06-10 02:59
【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。
[Abstract]:Objective to construct and identify rBCG-Rv2029c recombinant vaccine of Mycobacterium tuberculosis. Methods the Rv2029c antigen coding gene was amplified by PCR, then the Rv2029c and pMV261 plasmids were digested by double enzyme digestion, and then the digested products were ligated into rpMV261-2029c recombinant plasmid, and the plasmid was introduced into BCG to construct rBCG-Rv2029c recombinant vaccine by electroporation. Finally, the expressed recombinant protein was identified by SDS-PAGE and Western blotting. Results the 1 020bp Rv2029c gene was successfully amplified by PCR and inserted into pMV261 plasmid, and then the fusion gene was successfully introduced into BCG. It was confirmed by double enzyme digestion and gene comparison that the recombinant protein had immunogenicity by Western blotting after heat induction. Conclusion the recombinant live vaccine of Mycobacterium tuberculosis rBCG-Rv2029c was successfully constructed, which laid a foundation for the study of the immune mechanism of the recombinant vaccine.
【作者单位】： 南阳医学高等专科学校基础医学部;河南大学附属南阳南石医院;
【基金】：校自科NYYZ006~~
【分类号】：R392
[Abstract]:Objective to construct and identify rBCG-Rv2029c recombinant vaccine of Mycobacterium tuberculosis. Methods the Rv2029c antigen coding gene was amplified by PCR, then the Rv2029c and pMV261 plasmids were digested by double enzyme digestion, and then the digested products were ligated into rpMV261-2029c recombinant plasmid, and the plasmid was introduced into BCG to construct rBCG-Rv2029c recombinant vaccine by electroporation. Finally, the expressed recombinant protein was identified by SDS-PAGE and Western blotting. Results the 1 020bp Rv2029c gene was successfully amplified by PCR and inserted into pMV261 plasmid, and then the fusion gene was successfully introduced into BCG. It was confirmed by double enzyme digestion and gene comparison that the recombinant protein had immunogenicity by Western blotting after heat induction. Conclusion the recombinant live vaccine of Mycobacterium tuberculosis rBCG-Rv2029c was successfully constructed, which laid a foundation for the study of the immune mechanism of the recombinant vaccine.
【作者单位】： 南阳医学高等专科学校基础医学部;河南大学附属南阳南石医院;
【基金】：校自科NYYZ006~~
【分类号】：R392
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