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Nucleic Acids Research, 1985, Vol. 13, No. 1 59-71
© 1985

Articles

Promoter analysis of the phosphoenoipyruvate carboxylase gene of Escherichia coli

Katsura Izui, Tetsuya Miwa1, Masayuki Kajitani2,*, Nobuyuki Fujita3, Hisataka Sabe4, Akira Ishihama3,* and Hirohiko Katsuki5

Department of Chemistry, Faculty of Science, Institute for Virus Research, Kyoto University Kyoto 606, Japan *Department of Biochemistry, Institute for Virus Research, Kyoto University Kyoto 606, Japan

5To whom reprint request should be sent

Received October 8, 1984. Revised December 4, 1984. Accepted December 4, 1984.

In order to find the promoter region of phosphoenolpyruvate carboxylase [EC 4.1.1.31 [EC] ] 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 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.

1Present address: Central Research Laboratories, Ajinomoto Co., Inc., Kawasaki, Kanagawa-ken 210, Japan.

2Present address: Fundamental Research Laboratories, Toray Industries, Kamakura, Kanagawa-ken 248, Japan.

3Present address: National Institute of Genetics, Mishima, Shizuoka-ken 411, Japan.

4Present address: Institute for Virus Research, Kyoto University, Kyoto 606, Japan.


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