利用带有向外诱导型启动子的改良Tn7转座子对枯草芽孢杆菌进行体外诱变。
In vitro mutagenesis of Bacillus subtilis by using a modified Tn7 transposon with an outward-facing inducible promoter.
作者信息
Bordi Christophe, Butcher Bronwyn G, Shi Qiaojuan, Hachmann Anna-Barbara, Peters Joseph E, Helmann John D
机构信息
Department of Microbiology, Wing Hall, Cornell University, Ithaca, NY 14853-8101, USA.
出版信息
Appl Environ Microbiol. 2008 Jun;74(11):3419-25. doi: 10.1128/AEM.00476-08. Epub 2008 Apr 11.
Abstract
A Tn7 donor plasmid, pTn7SX, was constructed for use with the model gram-positive bacterium Bacillus subtilis. This new mini-Tn7, mTn7SX, contains a spectinomycin resistance cassette and an outward-facing, xylose-inducible promoter, thereby allowing for the regulated expression of genes downstream of the transposon. We demonstrate that mTn7SX inserts are obtained at a high frequency and occur randomly throughout the B. subtilis genome. The utility of this system was demonstrated by the selection of mutants with increased resistance to the antibiotic fosfomycin or duramycin.
摘要
构建了一种Tn7供体质粒pTn7SX，用于模式革兰氏阳性菌枯草芽孢杆菌。这种新的微型Tn7，即mTn7SX，包含一个壮观霉素抗性盒和一个向外的、木糖诱导型启动子，从而允许对转座子下游基因进行调控表达。我们证明，mTn7SX插入片段能以高频率获得，且随机出现在枯草芽孢杆菌基因组中。通过筛选对抗生素磷霉素或短杆菌肽耐药性增强的突变体，证明了该系统的实用性。
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