中文摘要：

目的：应用抑制性消减杂交技术，构建致肾盂肾炎大肠埃希菌132株（UPEC132）与已完成基因组测序的非致病性大肠埃希菌标准株MGl655的基因组DNA消减文库。方法：分别提取UPEC132与MG1655的基因组DNA，经RsaⅠ酶切成为大小200～800bp的片段，一部分与接头连接作为tester，和未与接头连接的酶切片段（driver）进行2次消减杂交及2次抑制性PCR后将产物与pMD18-T载体连接构建DNA消减文库，并转化给大肠埃希菌TOP10构建消减文库。结果：文库扩增后得到139个克隆，经PCR法快速测定，均得到200～800bp的插入片段。结论：所构建的基因组DNA消减文库为进一步筛选UPECl32中与致病相关基因奠定了基础。

英文摘要：

Objective： To constructed DNA library of uropathogenic E.coli132 （UPEC132） by suppression subtractive hybridization （SSH）. Methods： The genome DNA of UPEC132 and MG1655 were obtained （as tester and driver）, after Rsa I enzyme restriction, tester DNA were divided two groups to ligated to the specific adaptorl and adaptor2R. Then tester DNA were hybridized with driver DNA twice and underwent nested PCR twice. The PCR product was ligated with pMD18-T vector, transformed into E.coli TOP10, screened through the blue-white screening system. Results： 139 positive recombinant clones were confirmed by PCR method. Conclusion： The DNA subtractive library may provide a solid foundation for screening and cloning pathogenic genes of UPEC132.