一种用于洋葱伯克霍尔德菌及其他多重耐药革兰氏阴性菌基因操作的无标记缺失方法。
A markerless deletion method for genetic manipulation of Burkholderia cenocepacia and other multidrug-resistant gram-negative bacteria.
作者信息
Aubert Daniel F, Hamad Mohamad A, Valvano Miguel A
机构信息
Department of Microbiology and Immunology, Centre for Human Immunology, University of Western Ontario, London, ON, Canada, N6A 5C1.
出版信息
Methods Mol Biol. 2014;1197:311-27. doi: 10.1007/978-1-4939-1261-2_18.
Genetic manipulation of multidrug-resistant bacteria is often difficult and hinders progress in understanding their physiology and pathogenesis. This book chapter highlights advances in genetic manipulation of Burkholderia cenocepacia, which are also applicable to other members of the Burkholderia cepacia complex and multidrug-resistant gram-negative bacteria of other genera. The method detailed here is based on the I-SceI homing endonuclease system, which can be efficiently used for chromosomal integration, deletion, and genetic replacement. This system creates markerless mutations and insertions without leaving a genetic scar and thus can be reused successively to generate multiple modifications in the same strain.
对多重耐药细菌进行基因操作通常很困难，这阻碍了我们在了解其生理学和发病机制方面取得进展。本章重点介绍了洋葱伯克霍尔德菌基因操作方面的进展，这些进展也适用于洋葱伯克霍尔德菌复合体的其他成员以及其他属的多重耐药革兰氏阴性菌。此处详细介绍的方法基于I-SceI归巢内切酶系统，该系统可有效地用于染色体整合、缺失和基因替换。该系统产生无标记的突变和插入，不会留下遗传疤痕，因此可以连续重复使用，在同一菌株中产生多种修饰。
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