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Nucleic Acids Research, 1990, Vol. 18, No. 23 7049-7053
© 1990

Articles

Determination of the cis sequence involved in catabolite repression of the Bacillus subtilis gnt operon; implication of a consensus sequence in catabolite repression in the genus Bacillus

Yasuhiko Miwa and Yasutaro Fujita*

Department of Biotechnology, Faculty of Engineering, Fukuyama University Fukuyama 729-02, Japan

* To whom correspondence should be addressed

Received July 24, 1990. Revised November 5, 1990. Accepted November 5, 1990.

The mechanism underlying catabolite repression in Bacillus species remains unsolved. The gluconate (gnt) operon of Bacillus subtilis is one of the catabolic operons which is under catabolite repression. To identify the cis sequence involved in catabolite repression of the gnt operon, we performed deletion analysis of a DNA fragment carrying the gnt promoter and the gntR gene, which had been cloned into the promoter probe vector, pWP19. Deletion of the region upstream of the gnt promoter did not affect catabolite repression. Further deletion analysis of the gnt promoter and gntR coding region was carried out after restoration of promoter activity through the insertion of Internal constitutive promoters of the gnt operon before the gntR gene (P2 and P3). These deletions revealed that the cis sequence involved In catabolite repression of the gnt operon is located between nucleotlde positions +137 and +148. This DNA segment contains a sequence, ATTGAAAG, which may be implicated as a consensus sequence involved in catabolite repression in the genus Bacillus.


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