Localization and functional analysis of the regulated promoter from the Streptomyces glaucescens mel operon.
The transcription initiation site of the mel operon from Streptomyces glaucescens, determined by S1 mapping and primer elongation experiments, lies 32 to 34 bp upstream of the translation initiation codon of the first open reading frame. A total of 172 to 219 bp upstream of the transcription start point are necessary for a fully active and regulated mel promoter. Deletion analysis, gel retardation assays and DNAse I footprint experiments facilitated division of the promoter into three functional domains, which include the RNA polymerase recognition site up to nucleotides -33 to -42, the binding region of a protein of assumed regulatory function between nucleotides -65 and -93, and an upstream activator site, located between positions -158 and -219.
Geistlich
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Mikrobiologisches Institut, Eidgenössische Technische Hochschule, ETH-Zentrum, Zürich, Switzerland.
Irniger
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Hütter
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D004252:DNA Mutational Analysis; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009876:Operon; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D013302:Streptomyces / Q000235:genetics*; D014157:Transcription Factors; D014158:Transcription, Genetic; D014169:Transformation, Bacterial
D014157:Transcription Factors