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Nucleic Acids Research, 1983, Vol. 11, No. 11 3451-3465
© 1983

MOLECULAR BIOLOGY

Cloning and promoter analysis of the Escherichia coli adenylate cyclase gene

Hiroji Aiba1, Makoto Kawamukai2 and Akira Ishihama3

1Radioisotope Laboratory, Faculty of Medicine Kyoto 606, Japan 2Department of Agricultural Chemistry Kyoto 606, Japan 3Institute for Virus Reasearch, Kyoto University Kyoto 606, Japan

Received April 20, 1983. Accepted May 11, 1983.

The gene for adenylate cyclase of E. coli has been cloned in the plasmid pBR322. The Cya– strain transformed with the isolated plasmids produces significant amounts of adenylate cyclase and cAMP. Some of the Cya+plasmids were shown to direct the synthesis of a 85,000 dalton polypeptide in a cell-free system. The direction of transcription and the location of the cya promoter including the transcriptional start site were determined by an SI digestion method. DNA sequence around the promoter region indicates that a putative coding region for adenylate cyclase begins at +233. The 233 bp leader region could encode a potential small polypeptide containing 30 amino acids. Two probable CRP binding sites were found in the leader region, suggesting a negative control at the transcriptional level by CRP-cAMP.


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