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TOL is a broad-host-range plasmid

S Benson, J Shapiro

    Journal of Bacteriology
    |July 1, 1978
    PubMed
    Summary
    This summary is machine-generated.

    Researchers inserted the carbenicillin resistance element Tn401 into the TOL plasmid in Pseudomonas putida. The resulting hybrid plasmids conferred carbenicillin resistance in multiple bacterial species, but TOL-encoded functions were host-dependent.

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    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • The TOL plasmid is crucial for microbial degradation of aromatic hydrocarbons.
    • Gene transfer and expression across different bacterial species are fundamental to microbial evolution.
    • Carbenicillin resistance (Cbr) is a significant trait in bacterial pathogens.

    Purpose of the Study:

    • To investigate the stability and functional expression of hybrid TOL::Tn401 plasmids in different bacterial hosts.
    • To determine the host-specific expression of plasmid-encoded genes, including those for Cbr and m-toluate metabolism.

    Main Methods:

    • Isolation of Tn401 insertions into the TOL plasmid in Pseudomonas putida.
    • Introduction of hybrid TOL::Tn401 plasmids into Pseudomonas aeruginosa and Escherichia coli.

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  • Assessment of plasmid stability and expression of Cbr phenotype.
  • Evaluation of the ability to grow on m-toluate as an indicator of TOL function.

    • Main Results:

    • Hybrid TOL::Tn401 plasmids were successfully constructed and stably maintained in P. putida, P. aeruginosa, and E. coli.
    • The carbenicillin resistance (Cbr) phenotype was consistently expressed in all three bacterial hosts.
    • Replicative and conjugative functions of the TOL plasmid were expressed in both Pseudomonas and E. coli.
    • The ability to metabolize m-toluate, a key TOL plasmid function, was exclusively observed in Pseudomonas species.

    Conclusions:

    • Tn401 can be stably integrated into the TOL plasmid, creating hybrid plasmids with broad host compatibility for Cbr expression.
    • The expression of TOL-specific metabolic functions is restricted to the native host genus (Pseudomonas), highlighting host-dependent regulatory mechanisms.
    • This study provides insights into horizontal gene transfer and the functional divergence of plasmids in different bacterial genera.