1. ROMA: An in vitro approach to defining target genes for transcription regulators
- Author
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MacLellan, Shawn R., Eiamphungporn, Warawan, and Helmann, John D.
- Subjects
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GENETIC transcription regulation , *DNA microarrays , *RNA polymerases , *ANTISENSE DNA , *GENETIC regulation , *BACILLUS subtilis genetics - Abstract
Abstract: 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. [Copyright &y& Elsevier]
- Published
- 2009
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