Generation of Single-Copy Transposon Insertions in Clostridium perfringens by Electroporation of Phage Mu DNA Transposition Complexes
Generation of Single-Copy Transposon Insertions in Clostridium perfringens by Electroporation of Phage Mu DNA Transposition Complexes
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DOI：
10.1128/aem.02214-08
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发表时间：
2009-05-01
影响因子：
4.4
通讯作者：
Van Immerseel, F.
中科院分区：
文献类型：
--
作者：
Lanckriet, A.;Timbermont, L.;Van Immerseel, F.
转座子诱变是一种广泛用于鉴定与细菌毒力有关的基因的工具。到目前为止，产气荚膜梭菌中的转座子诱变仅限于使用基于 Tn916 的方法和实验室参考菌株。该系统主要在单个基因组中产生多个转座子插入，从而损害了其用于鉴定毒力基因的用途。目前的研究描述了产气荚膜梭菌转座子诱变的新方案，该方案基于噬菌体 Mu 转座系统。该方案已成功用于为实验室菌株和现场分离株生成单插入突变体库。因此，它可以用作大规模筛选鉴定产气荚膜梭菌毒力基因的工具。
Transposon mutagenesis is a tool that is widely used for the identification of genes involved in the virulence of bacteria. Until now, transposon mutagenesis in Clostridium perfringens has been restricted to the use of Tn916-based methods with laboratory reference strains. This system yields primarily multiple transposon insertions in a single genome, thus compromising its use for the identification of virulence genes. The current study describes a new protocol for transposon mutagenesis in C. perfringens, which is based on the bacteriophage Mu transposition system. The protocol was successfully used to generate a single-insertion mutant library both for a laboratory strain and for a field isolate. Thus, it can be used as a tool in large-scale screening to identify virulence genes of C. perfringens.