结核分枝杆菌新抗原Mtb8.4基因的克隆及真核表达载体的构建  

Construction and identification of the pcDNA3.1(+)-mtb8.4 eukaryotic expression plasmid

作　　者：李晖[1] 钟森[1] 任红[2] 史小玲[1] 

机构地区：[1]泸州医学院附属医院,四川泸州646000 [2]重庆医科大学病毒性肝炎研究所,重庆400010

出　　处：《四川医学》2003年第5期448-450,共3页Sichuan Medical Journal

基　　金：四川省青年科技基金资助 (批准号 :川青科基 [2 0 0 2 ] 1号 )

摘　　要：目的　克隆结核杆菌新抗原Mtb8.4基因 ,并构建真核表达载体pcDNA 3 .1( +) mtb8.4。方法　提取人结核杆菌H3 7Rv株的基因组作为模板 ,进行PCR扩增 ,获得mtb8.4目的基因后 ,与pcDNA 3 .1( +)载体进行连接重组。结果　pcDNA 3 .1( +) mtb8.4真核表达载体构建完成后 ,用限制性内切酶消化、PCR及DNA序列分析等多种方法进行鉴定 ,证实其构建成功。结论　pcDNA 3 .1( +) mtb8.4真核表达质粒的成功构建 ,为进一步研究该真核表达质粒的免疫保护效果及制备结核病pcDNA 3 .1( +) mtb8.4DNA疫苗奠定了基础。Objective To construct and identify the pcDNA3.1(+)-mtb8.4 eukaryotic expression plasmid.Methods Extracted DNA from M.tuberculosis was amplified by PCR and the target gene we got was cloned into the unique HindⅢand EcoR I cloning sites of pcDNA3.1(+).Results The accuracy of pc DNA 3.1(+)-mtb 8.4 plasmid construction was confirmed by a series of molecular biology techniques.Conclusion The construction of pcDNA 3.1(+)-mtb 8.4 may provide the possibilities of investigating immunogenicity of the recombinant plasmid and preparing a new tuberculosis vaccine.

关 键 词：结核杆菌 mdb8.4 PCR 基因克隆 

分 类 号：R392[医药卫生—免疫学]