Promoter analysis of the phosphoenolpyruvate carboxylase gene of Escherichia coli

Abstract

In order to find the promoter region of phosphoenolpyruvate carboxylase [EC 4.1.1.31] gene (ppc), in vitro transcription was performed using truncated DNA fragments as templates. Transcription mapping showed three promoters as candidates, but only one of them could be assigned to the promoter of ppc gene, considering the nucleotide sequence of its coding region (Fujita, N., Miwa, T., Ishijima, S., Izui, K. and Katsuki, H. (1984) J. Biochem. 95, 909-916). Nuclease S1 mapping showed that the in vivo and in vitro transcription initiation sites are identical and that the site lies 91 or 92 nucleotides upstream the translation initiation site. No alteration of the transcription initiation site was observed whether the cells were starved for an amino acid or grown on various carbon sources. The sequences of the -10 and -35 regions were fairly in accordance with the consensus sequences hitherto reported. Some features of the sequence around the promoter region were discussed.