副结核分枝杆菌hsp65基因的克隆及序列分析  

Cloning and Sequence Analysis of Mycobacterium paratuberculosis hsp65 gene

作　　者：徐凤宇[1] 胡玉庆[1] 曾范利[1] 姜秀云[2] 何昭阳[1] 

机构地区：[1]吉林农业大学动物科技学院,长春130118 [2]吉林农业大学生命科学学院,长春130118

出　　处：《吉林农业大学学报》2009年第2期204-207,共4页Journal of Jilin Agricultural University

基　　金：国家自然科学基金项目(30471285);吉林农业大学青年启动基金项目(2007001)

摘　　要：以副结核分枝杆菌C-2株基因组DNA为模板,以hsp65基因特异性引物进行PCR扩增,获得了1 626 bp的DNA片段。通过T-A克隆技术,将PCR产物克隆至pGEM-T载体。经鉴定,成功地构建出了重组质粒pGEM-T-hsp65。序列分析结果表明:该序列与GenBank中副结核分枝杆菌K-10株的同源性为99.2%。The genomic DNA was extracted from Mycobacterium paratuberculosis （MP） strain C - 2. The secreted protein hsp65 gene was amplified with a pair of specific primers using polymerase chain reaction （PCR）. PCR product was anapproximate 1 626 bp DNA segment. The clone vector pGEM-T-hsp65 was constructed successfully by the PCR product that was cloned into pGEM-T vector using T- A clone tech- nique, and the recombinant clone was identified by using complementation test, plasmid size test, re- strictional enzyme assay, plasmid PCR identification and recombinant plasmid sequence analysis. The analysis indicated that the hsp65 gene was very conservative in MP. These results could serve as a basis for further study on the usefulness of hsp65 gene and immunogenicity characteristic of hsp65 gene expression product.

关 键 词：副结核分枝杆菌 hsp65基因 克隆 序列分析 

分 类 号：S852.618[农业科学—基础兽医学] Q785[农业科学—兽医学]