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Following Cell-fate in E. coli After Infection by Phage Lambda
06:10

Following Cell-fate in E. coli After Infection by Phage Lambda

Published on: October 14, 2011

DNA rearrangements in a hybrid plasmid carrying the redB imm region of coliphage lambda

F Bernardi, A Bernardi

    Gene
    |April 1, 1980
    PubMed

    Insights

    The hybrid plasmid is unstable at 37°C, undergoing modifications like IS1 insertion, mutations, and large deletions. These changes suggest a potential site-specific recombination event in the lambda phage DNA fragment.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Virology

    Background:

    • Hybrid plasmids combining bacterial and phage DNA are tools for genetic studies.
    • Phage lambda DNA integration and maintenance are complex processes influenced by temperature.

    Purpose of the Study:

    • To investigate the stability and modifications of a pSC101-lambda hybrid plasmid at different temperatures.
    • To characterize the genetic alterations occurring in the plasmid at elevated temperatures.

    Main Methods:

    • Construction of a hybrid plasmid containing pSC101 and the redB-N-imm region of phage lambda cI857.
    • Culturing bacterial cells containing the plasmid at 30°C and 37°C.
    • Restriction mapping to analyze plasmid modifications.

    Main Results:

    • The hybrid plasmid is stable at 30°C but unstable at 37°C.
    • At 37°C, plasmids showed IS1 element insertion into the N gene, a mutation in the pL-oL site, and four large deletions (30-50%) of the lambda fragment.
    • Deletions exhibited a common endpoint on the pSC101 fragment and variable endpoints on the lambda fragment.

    Conclusions:

    • Temperature significantly impacts the stability of the pSC101-lambda hybrid plasmid.
    • The observed modifications, particularly deletions, suggest a temperature-dependent, site-specific recombination mechanism involving the lambda DNA fragment.

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