| 一株新分离的肠杆菌噬菌体的快速遗传鉴定及其宿主识别基因的快速克隆 |
| |
| 引用本文： | 姜焕焕,王盛,李存,刘大斌,于长明,安小平,米志强,陈建魁,童贻刚.一株新分离的肠杆菌噬菌体的快速遗传鉴定及其宿主识别基因的快速克隆[J].生物工程学报,2011,27(6):884-890. |
| |
| 作者姓名： | 姜焕焕 王盛 李存 刘大斌 于长明 安小平 米志强 陈建魁 童贻刚 |
| |
| 作者单位： | 1. 军事医学科学院微牛物流行病研究所,病原微生物生物安全国家重点实验室,北京100071
2. 军事医学科学院解放军307医院临床检验科,北京,100071 |
| |
| 基金项目： | 国家高技术研究发展计划 (863计划) (Nos. 2007AA02Z151, 2009AA02Z111) 资助。 |
| |
| 摘 要： | 以大肠杆菌8099为宿主自医院污水中分离出一株肠杆菌噬菌体IME08,其遗传物质经RNA酶、DNA酶处理证实其为DNA,用限制性内切酶处理该DNA证实其为双链DNA。利用随机引物PCR技术扩增并克隆该噬菌体基因组的随机片段,经测序后同源比对,判断该噬菌体是一株新的T4-like噬菌体。根据4株T4-like噬菌体 (T4,JS98,T2及K3) 宿主识别基因 (g37) 5'端的高度保守序列,采用随机PCR与巢式PCR结合的“基因组跳跃”策略快速克隆出了该噬菌体的宿主识别基因g37和g38。
|
| 关 键 词： | 肠杆菌噬菌体 T4噬菌体属 PCR 基因组跳跃 宿主识别基因 |
| 收稿时间： | 2010/11/24 0:00:00 |
Rapid genetic characterization of a novel Enterobacteria phage and determination of its host recognizing genes |
| |
| Huanhuan Jiang,Sheng Wang,Cun Li,Dabin Liu,Changming Yu,Xiaoping An,Zhiqiang Mi,Jiankui Chen and Yigang Tong.Rapid genetic characterization of a novel Enterobacteria phage and determination of its host recognizing genes[J].Chinese Journal of Biotechnology,2011,27(6):884-890. |
| |
| Authors: | Huanhuan Jiang Sheng Wang Cun Li Dabin Liu Changming Yu Xiaoping An Zhiqiang Mi Jiankui Chen and Yigang Tong |
| |
| Institution: | State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China;Clinical Laboratory of No. 307 Hospital, Academy of Military Medical Sciences, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China |
| |
| Abstract: | We isolated a novel Enterobacteria phage IME08 from hospital sewage, then confirmed it was a double-stranded DNA phage by digesting its genetic material with DNase I, RNase A and several restriction endonucleases respectively. BLAST results of random fragments generated by a random PCR cloning method revealed that it belonged to T4-like virus. We subsequently determined the host recognizing genes (g37 and g38) sequence with a PCR-based "genome jumping" protocol based on highly conserved region at 5' terminus of g37 from four other T4-like Bacteriophages (T4, JS98, T2 and K3). These molecular biological methods enabled us to readily characterize the bacteriophage and efficiently determine the sequence of the genes of interest based on very limited conserved sequence information. |
| |
| Keywords: | Enterobacteria phage T4-like virus PCR genome jumping host recognizing gene |
| 本文献已被 CNKI 万方数据 PubMed 等数据库收录！ |
| 点击此处可从《生物工程学报》浏览原始摘要信息 |
| 点击此处可从《生物工程学报》下载免费的PDF全文 |
|
