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Related Experiment Videos

Chloramphenicol-inducible gene expression in Bacillus subtilis.

E J Duvall, D M Williams, P S Lovett

    Gene
    |October 1, 1983
    PubMed
    Summary

    A cloned Bacillus pumilus cat gene shows chloramphenicol-inducible activity in Bacillus subtilis. This inducibility is linked to a specific DNA region, suggesting a mechanism involving RNA stem-loop destabilization by chloramphenicol.

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    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • The cat gene from Bacillus pumilus confers chloramphenicol acetyltransferase activity.
    • Understanding the regulation of this gene is crucial for antibiotic resistance studies.

    Purpose of the Study:

    • To investigate the molecular basis of chloramphenicol inducibility for the Bacillus pumilus cat gene in Bacillus subtilis.
    • To identify the specific DNA region responsible for this inducible trait.

    Main Methods:

    • Gene cloning and expression in Bacillus subtilis.
    • DNA sequencing and analysis of the cat gene region.
    • RNA secondary structure prediction.

    Main Results:

    • A 234-bp DNA region was identified as responsible for chloramphenicol inducibility.

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  • The cat ribosome-binding site was localized within a 40-bp segment containing inverted repeats.
  • Sequence analysis predicted a stem-loop RNA structure that sequesters the ribosome-binding site.

    • Conclusions:

    • Chloramphenicol inducibility of the cat gene is mediated by a specific DNA sequence.
    • The mechanism likely involves chloramphenicol-induced destabilization of an RNA stem-loop structure.
    • This provides insight into prokaryotic gene regulation and antibiotic resistance mechanisms.