真核表达质粒pcDNA3-hsp70的构建及鉴定  

CONSTRUCTION AND IDENTIFICATION OF THE EUKARYOTIC EXPRESSION PLASMID pcDNA3-hsp70

作　　者：李晖[1] 史小玲[1] 钟森[1] 邓存良[1] 张建军[1] 

机构地区：[1]泸州医学院附属医院感染病科,四川泸州646000

出　　处：《泸州医学院学报》2002年第4期281-284,共4页Journal of Luzhou Medical College

基　　金：国家自然科学基金资助课题 (39870 70 3)

摘　　要：目的 :构建并鉴定真核表达质粒pcDNA3-hsp70。方法 :用基因释放剂提取人结核杆菌H37Rv株的基因组作为模板 ,进行PCR扩增 ,获得hsp70目的基因后 ,与pcDNA3载体进行连接重组。结果 :pcDNA3-hsp70真核表达质粒构建完成后 ,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 :pcDNA3-hsp70真核表达质粒的成功构建 。Objecticves: To construct and identify the eukaryotic expression plasmid pcDNA3-hsp70. Methods: The genome extracted from M. Tuberculosis with genereleaser was amplified by PCR and the target gene hspto soobtained was linked to the unique HindⅢ and XbaⅠbinding sites of pcDNA3. Results: The accuracy of pcDNA3-hsp70 plasmid constructed was confirmed by a series of molecular biological techniques. Conclusion: The construction of pcDNA3-hsp70 provided the possibility of investigating the immunogenicity of the recombinant plasmid and preparing for a new tuberculosis vaccine.

关 键 词：结核杆菌 HSP70 基因释放剂 PCR 真核表达质粒 DNA疫苗 结核病 

分 类 号：R52[医药卫生—内科学]