【摘要】 目的:构建可表达融合蛋白IL-12p70-Ag85A的重组卡介苗,为结核病新疫苗的研制奠定基础。方法:以穿梭表达质粒pMV361为载体,构建重组质粒rpMV361-IL-12p70-Ag85A。将成功构建的重组质粒以电转化方式导入BCG基因组构建重组卡介苗rBCG-IL-12p70-Ag85A。通过抗生素筛选、重组卡介苗基因组PCR扩增及测序,以及western-blotting对重组卡介苗进行鉴定。结果:经鉴定,重组卡介苗成功构建,并可表达融合蛋白IL-12p70-Ag85A。结论:重组卡介苗rBCG-12p70-Ag85A构建成功,并可高效表达目的蛋白。更多还原

【Abstract】 Objective To construct a recombinant BCG expressing fusion protein of human IL-12p70-Ag85A,to lay the foundation for production of new tuberculosis vaccine.Methods Construct recombinant plasmid of rpMV361-IL-12p70-Ag85A using plasmid pMV361 as the carrier.Then,the recombinant plasmid was imported BCG genome by electric transformation mode to construct recombinant BCG vaccine of rBCG-IL-12p70-Ag85A.The recombinant BCG was certified by antibiotic selection,PCR amplification and sequencing of recombinant BCG genome and western-blotting.Results The recombinant BCG was certified to be constructed successfully expressing fusion protein of IL-12p70-Ag85A.Conclusion Recombinant rBCG-IL-12p70-Ag85A is constructed successfully,and can highly express the target protein.更多还原