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Identification of nucleotides critical for activity of the Pseudomonas putida catBC promoter

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Summary

Pseudomonas putida utilizes the catBC operon, which encodes cis,cis-muconate lactonizing enzyme I (MLEI; EC 5.5.1.1) and muconolactone isomerase (MI; EC 5.3.3.4), for growth on benzoate as a sole carbon source. This operon is positively regulated, and the promoter is located 64 bp upstream of the catB translational start site. Using site-specific mutagenesis, we identified nucleotides that influenced the induction of this promoter. Promoter activity was monitored with the promoter probe vector pKT240. Transcription of mRNA from mutant promoters was determined by primer extension mapping. Comparison of the initiation start site of mutant promoters with that of the wild-type promoter identified a single functional promoter.

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Abbreviations

aph :

aminoglycoside phosphotransferase gene

bp :

base pairs

MI:

muconolactone isomerase

MLEI:

cis,cis-muconate lactonizing enzyme I

BSM:

basal salts medium

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Author notes

  1. Teri L. Aldrich

    Present address: Department of Genetics, University of Washington, 98195, Seattle, WA, USA

Authors and Affiliations

  1. Department of Microbiology and Immunology, University of Illinois College of Medicine, 60612, Chicago, IL, USA

    Teri L. Aldrich, Randi Kubrick Rothmel & A. M. Chakrabarty

Authors
  1. Teri L. Aldrich
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  2. Randi Kubrick Rothmel
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  3. A. M. Chakrabarty
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Communicated by C.P. Hollenberg

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Aldrich, T.L., Rothmel, R.K. & Chakrabarty, A.M. Identification of nucleotides critical for activity of the Pseudomonas putida catBC promoter. Molec Gen Genet 218, 266–271 (1989). https://doi.org/10.1007/BF00331277

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  • DOI: https://doi.org/10.1007/BF00331277

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