摘要
目的 通过对肺炎克雷伯菌KF3质粒DNA全序列测定,从基因组水平研究质粒DNA的结构、功能基因和与宿主菌耐药相关性.方法 碱裂解法提取质粒DNA,构建质粒DNA文库并测序.采用Phred/Phrap/Consed软件包进行序列拼接,Glimmer软件预测开放阅读框架（ORF）及功能分析.结果 构建包含3个质粒DNA的pUC18文库和Fosmid文库,测序获得3个质粒全序列.功能注释分析发现3个质粒均为可接合转移质粒,编码大量耐药相关基因.结论 肺炎克雷伯菌KF3的3个质粒都是可接合转移质粒,将耐药基因在细菌间进行水平转移,造成了耐药菌的播散.
Objective To study the structures of the plasmids of Klebsiella pneumoniae KF3 at the genome metagenome level througth with whole plasmid DNA sequencing, to analyze the functional genes carried by plasmid and to identify the correlation of resistance and pathogenicity between the plasmids and the host strains. Methods The alkaline lysis method was used to extract plasmids. We constructed the small insert pUC18 library and the large insert Forsmid library, sequenced and used the Phred / Phrap / Consed package to assemble these sequences and gained a complete sequence. The open reading frame（ORFs） were predicted by the Glimmer software and annotated, analyzed the functions of these genes. Results We successfully constructed the pUC18 library and the Fosmid libraries for the plasmid DNA and obtained three circular double-stranded DNA plasmids： pKF3-70 （69 477 bp）, pKF3-90 （91 327 bp） and pKF3-147 （ 147 416 bp）. There were drug resistant genes, conjugative transfer genes and mobile DNA elements identified on three plasmids. Conclusion The three plasmids of KF3 could be transferred among different strains. It would lead to the dissemination of the resistant genes.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2010年第10期892-896,共5页
Chinese Journal of Microbiology and Immunology
基金
基金项目：浙江省自然科学基金项目（Y2081093）
浙江省科技计划项目（2008C23074）：温州市科技计划项目（H20100022）
关键词
质粒
耐药基因
接合转移基因
肺炎克雷伯菌
Plasmids
Drug resistant genes
Conjugative transfer genes
Klebsiella pneumoniae
