ROMA: an in vitro approach to defining target genes for transcription regulators.
ROMA: an in vitro approach to defining target genes for transcription regulators.
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ROMA：一种定义转录调节因子靶基因的体外方法。
DOI：
10.1016/j.ymeth.2008.10.009
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发表时间：
2009
期刊：
影响因子：
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通讯作者：
Helmann,JohnD
中科院分区：
文献类型：
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作者：
Maclellan,ShawnR;Eiamphungporn,Warawan;Helmann,JohnD
我们描述了一种基于体外转录的方法，称为ROMA（径流转录微阵列分析），用于全基因组分析由sigma因子和其他转录调节因子调节的转录。ROMA使用纯化的RNA聚合酶（含或不含调节蛋白）来监测基因组DNA模板的转录产物。转录后的RNA被转化为cDNA，并与基因阵列杂交，从而鉴定出被调节因子特异性激活的基因。我们讨论了在枯草芽孢杆菌中使用ROMA来定义sigma因子调控及其在不同物种中广泛应用于定义其他转录调控因子的调控。
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.