In vitro transcriptional analysis of the cytochrome P-450cam hydroxylase operon.
To characterize the promoters of cytochrome P-450cam hydroxylase operon (camDCAB) and the repressor gene (camR), in vitro run-off transcription assays were performed using RNA polymerase (RNAP) holoenzyme reconstituted with the core enzyme and the sigma70 protein of Pseudomonas aeruginosa. Both the mRNAs of camDCAB and camR were accurately transcribed from the respective promoter by the reconstituted RNAP holoenzyme. Both the transcriptions were repressed by CamR protein and the repressions were suppressed by D-camphor, consistent with the regulation in P. putida. These results suggest that the RNA polymerase containing sigma70 recognizes the promoter of camDCAB as well as that of camR.
Aramaki
H
H
Department of Molecular Biology, Daiichi College of Pharmaceutical Sciences, Fukuoka, Japan.
Sagara
Y
Y
Fujita
M
M
D001426:Bacterial Proteins; D001483:Base Sequence; D019475:Camphor 5-Monooxygenase / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D008969:Molecular Sequence Data; D009876:Operon / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics*; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D019475:Camphor 5-Monooxygenase