摘要
目的:应用信号标签突变技术(Signature tagged mutagenesis,STM)构建幽门螺杆菌突变体文库重组质粒。方法:采用平末端内切酶随机酶切幽门螺杆菌基因组DNA,并回收300～500bp片段(记作Fr),基因重组技术构建重组质粒pID700-Fr,电转化E.coliDH5α筛选阳性克隆,提取重组质粒酶切鉴定。结果:成功构建了1200个幽门螺杆菌STM文库重组质粒。结论:STM技术可用于幽门螺杆菌致病以及耐药相关基因的筛选,为幽门螺杆菌致病及耐药机理的研究奠定了基础。也将为幽门螺杆菌的治疗提供新的药物靶点。
Objective：To construct recombinant plasmids of Helicobacter pylori mutant library with Signature tagged mutagenesis （STM）. Methods：Total DNA of Helicobacter pylori was digested at random with blunt-end restriction enzyme, and the 300-500bp DNA fragments（Fr） after digestion were recovered. Recombinant vectors pID700-Fr were constructed, and then were electroporated into E.coli DH5α . Positive clones containing pID700-Fr were obtained by selection from chloramphenicol-resistance plates, recombinant plasmids pID700-Fr were identified by BamH I /Xho I digestion.Results：This study has successfully constructed 1200 recombinant plasmids of the Helicobacter pylori for the mutant library.Conclusions：STM technology might be used as genetic screening method of virulence and drug-resistance genes, which laid a foundation for study on the pathogenesis and drug resistance mechanism, and for further study on new drug targets of the H.pylori＇ s therapy.
出处
《现代生物医学进展》
CAS
2010年第5期855-857,共3页
Progress in Modern Biomedicine
关键词
幽门螺杆菌
信号标签诱变技术
重组质粒
Helicobacter pylori
Signature-tagged mutagenesis
Recombinant plasmid
