PMID: 6408264Jun 25, 1983Paper

Deletion mutagenesis of the Escherichia coli galactose operon promoter region

Journal of Molecular Biology
S BusbyH Buc

Abstract

Using recombinant DNA technology we have created a series of progressively longer deletions both upstream and downstream from the Escherichia coli galactose operon regulatory region. The effects of these lesions on expression of the two overlapping galactose promoters have been quantitated after DNA fragments carrying these deletions were cloned in a plasmid vector, in which the beta-galactosidase gene could be expressed from the truncated galactose regulatory region. The results allow us to determine which sequences are necessary for the activity of the two promoters. Our results show that for the P1 promoter, which is controlled by the cyclic AMP-cyclic AMP receptor protein complex (cAMP-CRP), the sequence necessary for full activity starts 56 base-pairs upstream from the transcription initiation point. In contrast, for the P2 promoter, which functions in the absence of cAMP-CRP, the crucial sequence extends to only 39 base-pairs upstream from the transcription start. Deletions that cut into these sequences cause reductions in promoter strength, although some promoter activity is observed even when the "-35 region" of both P2 and P1 are deleted. Analysis of deletions originating downstream from the regulatory region shows tha...Continue Reading

References

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Citations

Apr 5, 1985·Journal of Molecular Biology·M DreyfusS Busby
Jan 1, 1985·Gene·T A VölkerT A Bickle
Oct 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·K ForsmanB E Uhlin
Dec 17, 2008·Infection and Immunity·Maria D Bodero, George P Munson
Aug 4, 2006·Journal of Bacteriology·Carlos BalsalobreBernt Eric Uhlin
Sep 20, 2011·Journal of Bacteriology·Sheetal UppalNarendra Jawali
Oct 29, 2000·Journal of Bacteriology·M WestermarkB E Uhlin
Oct 4, 2003·Journal of Bacteriology·Anne M L BarnardStephen J W Busby
Mar 5, 2004·Journal of Bacteriology·Neil R WybornJeffrey Green
Sep 26, 2007·Molecular Microbiology·James K LithgowJeffrey Green
Sep 28, 2016·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Bandar SendyJack A Bryant
Feb 11, 2014·Journal of Bacteriology·Sheetal UppalNarendra Jawali

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