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目的:构建一种新型的、含结核分枝杆菌(M.S)抗原PPE68的耻垢分枝杆菌载体疫苗。方法:以结核分枝杆菌标准株H37Rv的基因组为模板,PCR方法获得Rv3873基因,然后通过克隆构建获得真核表达质粒pVAX-1-Rv3873,最后通过电转化的方法将pVAX-1-Rv3873真核表达质粒转化至耻垢分枝杆菌的感受态细胞中获得重组的耻垢分枝杆菌载体疫苗rM.S-PPE68。结果:成功构建了含结核分枝杆菌抗原PPE68的耻垢分枝杆菌载体疫苗。结论:该疫苗的构建为结核病的预防,尤其在高免疫保护作用的疫苗开发方面,提供了新的研究策略。
Objective: To construct a new M. smegmatis vector vaccine containing Mycobacterium tuberculosis (M.S) antigen PPE68. Methods: The Rv3873 gene was obtained by PCR using the genome of Mycobacterium tuberculosis H37Rv as a template. The eukaryotic expression plasmid pVAX-1-Rv3873 was obtained by cloning. Finally, the eukaryotic expression vector pVAX-1-Rv3873 Expression plasmid was transformed into M. smegmatis competent cells to obtain a recombinant M. smegmatis vector vaccine rM.S-PPE68. Results: Mycobacterium smegmatis vector vaccine containing Mycobacterium tuberculosis antigen PPE68 was successfully constructed. Conclusion: The construction of this vaccine provides a new strategy for the prevention of tuberculosis, especially in vaccine development with high immunoprotection.