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Related Experiment Videos

Deletion mutants from R plasmids with increased copy number.

Y Ike, H Hashimoto, S Mitsuhashi

    Microbiology and Immunology
    |January 1, 1979
    PubMed
    Summary

    Researchers studied R mutants of the Rms 201 plasmid, observing altered drug resistance and copy numbers. They mapped the plasmid

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    Area of Science:

    • Molecular Biology
    • Microbiology
    • Genetics

    Background:

    • The conjugative plasmid Rms 201 confers resistance to ampicillin, tetracycline, chloramphenicol, streptomycin, and sulfonamides.
    • R mutants Rms 201-12 and Rms 201-46 exhibit increased copy numbers and altered drug resistance profiles compared to the parent plasmid.

    Purpose of the Study:

    • To characterize R mutants of the Rms 201 plasmid.
    • To determine the gene order and regulatory elements of the Rms 201 plasmid and its mutants.

    Main Methods:

    • Isolation and characterization of tetracycline (tet) gene-deleted mutants.
    • Prolonged culture of R mutant cells to generate various deletion mutants.
    • Comparison of genetic markers from deletion mutants to establish a circular gene order.

    Main Results:

    • Mutants showed increased resistance to ampicillin, streptomycin, and chloramphenicol, but decreased resistance to tetracycline.
    • Tet-deleted R mutants were isolated from cells grown on high tetracycline concentrations.
    • A circular gene order for Rms 201 was proposed based on deletion mutant analysis.
    • Copy number regulation genes are closely linked to the rep gene.
    • Entry exclusion genes were found to be co-lost with the tra region.

    Conclusions:

    • The study elucidated the genetic organization of the Rms 201 plasmid and its mutants.
    • Key genes regulating plasmid copy number and entry exclusion were identified in relation to their genetic location.

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