Genomic studies with Escheria coli MelR protein: applications of chromatin immunoprecipitation and microarrays

David Grainger, Timothy Overton, JT Wade, E Tamai, Jonathan Hobman, Chrystala Constantinidou, Stephen Busby

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

Escherichia coli MelR protein is a transcription activator that is essential for melibiose-dependent expression of the melAB genes. We have used chromatin immunoprecipitation to study the binding of MelR and RNA polymerase to the melAB promoter in vivo. Our results show that MelR is associated with promoter DNA, both in the absence and presence of the inducer melibiose. In contrast, RNA polymerase is recruited to the melAB promoter only in the presence of inducer. The MelR DK261 positive control mutant binds to the melAB promoter but cannot recruit RNA polymerase. Further analysis of immunoprecipitated DNA, by using an Affymetrix GeneChip array, showed that the melAB promoter is the major, if not the sole, target in E. coli for MelR. This was confirmed by a transcriptomics experiment to analyze RNA in cells either with or without melR.
Original languageEnglish
Pages (from-to)6938-6943
Number of pages6
JournalJournal of Bacteriology
Volume186
DOIs
Publication statusPublished - 1 Jan 2004

Fingerprint

Dive into the research topics of 'Genomic studies with Escheria coli MelR protein: applications of chromatin immunoprecipitation and microarrays'. Together they form a unique fingerprint.

Cite this