嗜麦芽寡养单胞菌D2株基因组文库的构建及部分克隆的分析  

The genomic library construction and partial clone sequences analysis of stenotrophomonas maltophilia D2

作　　者：杨丽[1] 闫志勇[1] 王斌[1] 辛晓妮[1] 宋旭霞[1] 赵巍[1] 钱冬萌[1] 苏洁[1] 

机构地区：[1]青岛大学医学院微生物学教研室,青岛266071

出　　处：《医学研究生学报》2010年第4期369-371,共3页Journal of Medical Postgraduates

基　　金：国家"973"计划重大基础研究前期研究专项基金(2004CCA02400);山东省科技公关基金(2007GG3WZ05009);青岛市自然科学基金(07-2-3-8-jch)

摘　　要：目的自双齿围沙蚕消化道内分离出的嗜麦芽寡养单胞菌D2株经前期研究发现其具有明显胞外分泌可溶性纤溶酶、蛋白酶等活性。文中构建嗜麦芽寡养单胞菌D2株基因组文库,了解其基因背景。方法用Sau3AⅠ酶切D2株基因组DNA,回收0.2~2kb的DNA片段,连接至pMD18-T质粒载体,转化大肠杆菌JM109,在含有Amp的LB平板上筛选重组子,酶切鉴定后建库保存;随机挑取15个酶切鉴定正确的阳性克隆测序,在网络数据库中进行BLASTX分析。结果共得到转化子约1124个,符合建库要求;经测序证实获得了D2株碱性磷酸酶、AMP-依赖性合成酶、荧光素样单加氧酶等蛋白的结构基因序列。结论成功构建了嗜麦芽寡养单胞菌D2株的基因组文库,为进一步了解该菌的背景及筛选其功能基因奠定基础。Objective We obtained a Stenotrophomonas maltophilia D2 from the digestive tract of Perinereis aibuhitensis Grube which secreted fibrinogenase and protease. To construct a genomic library of Stenotrophomonas maltophilia D2 and to investigate the background of it. Methods Genomic DNA was digested with Sau3A Ⅰ, 0.2～2kb fragments were recovered, and inserted into pMD18-T vector, and then transformed into JM109. The recombinants were screened on the LB plate with Amp. All of the positive clones were stored, Resistant clones were randomly selected and then sequenced .The sequences were searched in NCBI using BLASTX tool. Results 1124 recombinants was obtained. The gene of alkaline phosphatase, AMP-dependent synthetase and ligase, and Luciferase-like monooxygenase were obtained. Conclusion A genomic library of Stenotrophomonas maltophilia D2 is successfully constructed. The results help us to know more about the background of the Stenotrophomonas maltophilia genome and to sieve its functional gene.

关 键 词：嗜麦芽寡养单胞菌 基因组文库 克隆 序列分析 

分 类 号：Q939[生物学—微生物学]