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.
出处
《四川医学》
CAS
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 )
