Genomic studies with Escherichia coli MelR protein:: Applications of chromatin immunoprecipitation and microarrays

被引:84
作者
Grainger, DC [1 ]
Overton, TW
Reppas, N
Wade, JT
Tamai, E
Hobman, JL
Constantinidou, C
Struhl, K
Church, G
Busby, SJW
机构
[1] Univ Birmingham, Sch Biosci, Birmingham B15 2TT, W Midlands, England
[2] Kagawa Univ, Fac Med, Dept Microbiol, Kagawa, Japan
[3] Harvard Univ, Sch Med, Dept Biol Chem & Mol Pharmacol, Boston, MA 02115 USA
[4] Harvard Univ, Sch Med, Dept Genet, Boston, MA 02115 USA
关键词
D O I
10.1128/JB.186.20.6938-6943.2004
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页码:6938 / 6943
页数:6
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