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Related Experiment Videos

Regular segregation of composite plasmid Rms201

Y Ike, H Hashimoto, S Mitsuhashi

    Journal of Bacteriology
    |November 1, 1981
    PubMed
    Summary

    New copy number mutants of the Rms201 plasmid were identified. These mutants demonstrate that Rms201 plasmid segregation occurs through a regular partitioning mechanism, impacting cell populations under temperature stress.

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    Area of Science:

    • Molecular Biology
    • Microbiology
    • Genetics

    Background:

    • The conjugative plasmid Rms201 is essential for bacterial conjugation.
    • Understanding plasmid replication and segregation is crucial for genetic stability.
    • Temperature-sensitive mutants provide tools to study essential genetic processes.

    Purpose of the Study:

    • To isolate and characterize new copy number mutants of the Rms201 plasmid.
    • To investigate the mechanism of Rms201 plasmid segregation.
    • To determine the impact of temperature shifts on plasmid replication and stability.

    Main Methods:

    • Isolation of temperature-sensitive copy number mutants (Rms201ts15, Rms201ts16) from a parent mutant (Rms201ts14).
    • Culturing bacterial strains (ML1410) with different Rms201 mutants at permissive (30°C) and restrictive (42°C) temperatures.
    • Quantifying plasmid copy number per chromosome and monitoring the appearance of antibiotic-sensitive segregants over generations.

    Main Results:

    • Mutants Rms201ts15 and Rms201ts16 exhibited increased plasmid copy numbers (7.4 and 20, respectively) compared to the parent mutant (2.2) at 30°C.
    • Plasmid DNA synthesis ceased in all mutants upon temperature shift to 42°C.
    • Antibiotic-sensitive segregants appeared after lag times ranging from 2.5 to 6.8 generations, with plasmid-positive cells decreasing by ~50% per generation post-lag.

    Conclusions:

    • The Rms201 plasmid utilizes a regular segregation (partition) mechanism for distribution into daughter cells.
    • The identified mutants provide insights into the regulation of plasmid copy number and segregation.
    • Temperature-sensitive mutations disrupt Rms201 replication and lead to loss of plasmid-bearing cells through regular segregation.

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