Maclellan Shawn R, Eiamphungporn Warawan, Helmann John D
Department of Microbiology, Cornell University, 327 Wing Hall, Ithaca, NY 14853-8101, USA.
Methods. 2009 Jan;47(1):73-7. doi: 10.1016/j.ymeth.2008.10.009. Epub 2008 Oct 21.
We describe an in vitro transcription-based method called ROMA (run-off transcription-microarray analysis) for the genome-wide analysis of transcription regulated by sigma factors and other transcriptional regulators. ROMA uses purified RNA polymerase with and without a regulatory protein to monitor products of transcription from a genomic DNA template. Transcribed RNA is converted to cDNA and hybridized to gene arrays allowing for the identification of genes that are specifically activated by the regulator. We discuss the use of ROMA to define sigma factor regulons in Bacillus subtilis and its broad application to defining regulons for other transcriptional regulators in various species.
我们描述了一种基于体外转录的方法，称为ROMA（ runoff转录-微阵列分析），用于全基因组分析由σ因子和其他转录调节因子调控的转录。ROMA使用纯化的RNA聚合酶，分别加入和不加入调节蛋白，以监测来自基因组DNA模板的转录产物。转录的RNA被转化为cDNA并与基因阵列杂交，从而能够鉴定被调节因子特异性激活的基因。我们讨论了ROMA在定义枯草芽孢杆菌中σ因子调控子方面的应用，以及它在定义各种物种中其他转录调节因子调控子方面的广泛应用。
