Kim Jeong Nam, Youm Gun Woong, Kwon Young Min
Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR 72701, USA.
Curr Microbiol. 2008 Oct;57(4):391-4. doi: 10.1007/s00284-008-9225-6. Epub 2008 Aug 14.
TnAraOut is a mariner-based transposon containing an arabinose-inducible promoter P(BAD) facing outward. TnAraOut mutagenesis previously used to identify essential genes in Vibrio cholerae can also be used to identify in vitro essential genes in Salmonella enteritidis. A mutant screen was conducted based on the assumption that a mutant-harboring TnAraOut insertion in the promoter region of an essential gene should exhibit arabinose-dependent growth phenotype. Among five isolated mutants with such growth phenotype, DNA sequencing revealed that two of them have insertions in the upstream region of atpI and the coding region of yigP gene such that P(BAD) promoter drives the expression of the downstream gene(s). Growth assay showed that the growth defects of these two mutants were fully restored by arabinose induction.
TnAraOut是一种基于水手座的转座子,含有一个向外的阿拉伯糖诱导型启动子P(BAD)。先前用于鉴定霍乱弧菌中必需基因的TnAraOut诱变也可用于鉴定肠炎沙门氏菌中的体外必需基因。基于这样的假设进行了突变体筛选:在必需基因启动子区域插入TnAraOut的突变体应表现出阿拉伯糖依赖性生长表型。在五个具有这种生长表型的分离突变体中,DNA测序显示其中两个在atpI上游区域和yigP基因编码区域有插入,使得P(BAD)启动子驱动下游基因的表达。生长试验表明,这两个突变体的生长缺陷通过阿拉伯糖诱导得以完全恢复。