从鳗弧菌中克隆出一个类插入序列元件并在大肠杆菌中对其进行功能鉴定

Molecular cloning of an insertion sequence-like element from Vibrio anguillarum and its functional identification in E. coli.

作者信息

Liang Na, Wu Haizhen, Ye Jiang, Xu Peng, Zhang Yuanxing, Zhang Huizhan

机构信息

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.

出版信息

Biotechnol Lett. 2007 Dec;29(12):1951-7. doi: 10.1007/s10529-007-9485-4. Epub 2007 Sep 7.

DOI:10.1007/s10529-007-9485-4

PMID:17823776

Abstract

The tdh (thermostable direct hemolysin) gene occurs in some strains of Vibrio species. All tdh genes are flanked by insertion sequence-like elements (ISV). All previous attempts have failed to detect transposition of these ISVs. In this work, we have built a transposition detection system in E. coli and succeeded in detecting the transposition of an insertion sequence-like element at a frequency of Km(r) mutants of 7.2 x 10(-6). A specific flanking sequence (5'-Py-Pu-3') was found on either side of the target duplication.

摘要

tdh（耐热直接溶血素）基因存在于某些弧菌属菌株中。所有tdh基因都侧翼有类插入序列元件（ISV）。此前所有检测这些ISV转座的尝试均未成功。在本研究中，我们构建了一个大肠杆菌转座检测系统，并成功以7.2×10⁻⁶的卡那霉素抗性（Km(r)）突变体频率检测到一个类插入序列元件的转座。在靶标重复序列两侧均发现了一个特定的侧翼序列（5'-Py-Pu-3'）。