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Plasmid-determined bleomycin resistance in Staphylococcus aureus.

D Semon, N R Movva, T F Smith

    Plasmid
    |January 1, 1987
    PubMed
    Summary
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    Researchers cloned and sequenced a fragment from Staphylococcus aureus plasmid pUB110, identifying a gene conferring bleomycin resistance. This gene, when expressed in E. coli, produced a 14,000-Da protein and conferred high-level resistance.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • Antibiotic resistance is a growing global health concern.
    • Plasmids are extrachromosomal DNA elements that can carry resistance genes.
    • Staphylococcus aureus is a significant human pathogen known for developing antibiotic resistance.

    Purpose of the Study:

    • To clone and sequence the bleomycin resistance gene from Staphylococcus aureus plasmid pUB110.
    • To characterize the expressed protein responsible for bleomycin resistance.
    • To compare the bleomycin resistance determinant with other known resistance elements.

    Main Methods:

    • Cloning and sequencing of a 1580-bp fragment from pUB110.
    • Subcloning of an open reading frame into expression vectors.

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  • Expression in Escherichia coli under the control of the lambda PL promoter.
  • Detection of the expressed protein using gel electrophoresis.

    • Main Results:

    • A 1580-bp fragment of pUB110 conferring bleomycin resistance was identified and sequenced.
    • A 405-bp open reading frame within this fragment was found to encode the resistance.
    • High-level bleomycin resistance was achieved in E. coli upon induction.
    • A protein of approximately 14,000 Da was detected, corresponding to the open reading frame.
    • Limited sequence relatedness was observed between the pUB110 determinant and Tn5.

    Conclusions:

    • The cloned DNA fragment from pUB110 contains a functional gene for bleomycin resistance.
    • The expressed protein is likely responsible for mediating resistance to bleomycin.
    • The findings contribute to understanding antibiotic resistance mechanisms in Gram-positive bacteria.