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Related Experiment Videos

Endpoint distribution for deletions into imm lambda region forming p lambda CM replicons: phage lambda gene rex

L A MacHattie

    Gene
    |January 1, 1985
    PubMed
    Summary

    The rexB gene product in lambda-derived plasmids is crucial for their establishment in new host cells. Loss of the rex gene leads to temperature-sensitive plasmid establishment, impacting early transduction stages.

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    Area of Science:

    • Molecular Biology
    • Bacteriology
    • Genetics

    Background:

    • Lambda-derived plasmids, specifically the p lambda CM family, confer chloramphenicol resistance (CmR).
    • Plasmid establishment is typically facilitated by the rexB gene product.
    • Deletions within the lambda immunity region can lead to p lambda CM formation.

    Purpose of the Study:

    • To investigate the role of the rex gene in the establishment of p lambda CM plasmids.
    • To characterize the phenotypic differences in plasmid establishment based on rex gene status.
    • To elucidate the specific function of Rex during early plasmid transduction stages.

    Main Methods:

    • Utilizing a 'groplaque' assay to distinguish between different classes of p lambda CM isolates based on growth characteristics.

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  • Employing temperature shifts (37°C to 32°C) to observe CmR growth patterns.
  • Conducting T4rII phage exclusion tests to identify Rex-deficient variants.

    • Main Results:

    • Two classes of p lambda CM isolates were identified: 'donut' groplaques (hollow-centered) and 'solid' groplaques.
    • 'Donut' groplaques resulted from deletions extending into or beyond the rexB gene, appearing only after a temperature shift.
    • 'Solid' groplaques, observed before the temperature shift, were associated with deletions not affecting rexB.
    • A Rex-deficient variant exhibited temperature-sensitive (ts) plasmid establishment but unaffected plasmid maintenance.

    Conclusions:

    • The rex gene is essential for the temperature-independent establishment of p lambda CM plasmids.
    • Rex likely functions in the early stages of transduction, potentially by aiding plasmid replication or inhibiting host replication.
    • This mechanism ensures the plasmid replicon achieves sufficient copy number for stable transmission.