PMID:8809031
| Citation |
Keen, J, Williams, J and Busby, S (1996) Location of essential sequence elements at the Escherichia coli melAB promoter. Biochem. J. 318 ( Pt 2):443-9 |
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| Abstract |
The Escherichia coli melAB promoter has been cloned on a short DNA fragment and subjected to deletion mutagenesis, random mutagenesis and site-directed mutagenesis. In previous work we had shown that expression from the melAB promoter is triggered by melibiose and that this requires the MelR transcription activator. Melibiose-dependent expression is suppressed by deletions that remove both DNA-binding sites for MelR and by point mutations in the -10 hexamer, the -35 hexamer and the region just upstream of the -35 hexamer. The point mutations identify promoter elements that are essential for triggering the melAB promoter. The importance of these elements was confirmed by site-directed mutagenesis. The results show that the organization of the melAB promoter is fundamentally different from the organization of other bacterial promoters controlled by homologues of MelR. |
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| Keywords |
Base Sequence; Binding Sites; Cloning, Molecular; DNA-Binding Proteins/metabolism; Deoxyribonuclease EcoRI; Deoxyribonuclease HindIII; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli Proteins; Molecular Sequence Data; Mutagenesis, Site-Directed; Plasmids; Point Mutation; Polymerase Chain Reaction; Promoter Regions, Genetic; Recombinant Fusion Proteins/biosynthesis; Restriction Mapping; Sequence Deletion; Trans-Activators/metabolism; beta-Galactosidase/biosynthesis |
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