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Related Experiment Videos

[Thermosensitive vector derived from RP1 plasmid for detection of transposable elements].

V N Danilevich, F A Amosenko, I Ia Volozhantseva

    Molekuliarnaia Biologiia
    |July 1, 1982
    PubMed
    Summary

    Transposable DNA elements in E. coli K12 mediate plasmid integration and Hfr formation. A new element, IS111, was discovered, alongside IS1- and IS5-like elements, aiding in understanding bacterial recombination.

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    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • Investigates the role of transposable DNA elements in bacterial chromosome integration.
    • Focuses on the RP1 plasmid and its derivatives in E. coli K12.

    Purpose of the Study:

    • To elucidate the mechanism of integrative recombination of the RP1 plasmid mediated by E. coli K12 IS-elements.
    • To identify and characterize novel insertion sequence (IS) elements involved in this process.

    Main Methods:

    • Utilized a temperature-sensitive plasmid RP1ts12, a derivative of RP1 with a mutated transposon Tnl.
    • Employed restriction and heteroduplex analysis for identification and characterization of IS elements in recombinant plasmids.
    • Assessed the activity of IS elements in recA-dependent integrative recombination.

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    Main Results:

    • Demonstrated that integration of RP1 into the E. coli K12 chromosome and Hfr formation can be mediated by chromosomal IS-elements.
    • Discovered a new IS-element, IS111 (approx. 1500bp), in E. coli K12, and identified IS1-like and IS5-like elements in RP1ts12 recombinants.
    • Concluded that E. coli K12 chromosomal IS-elements are absent in the RP1 plasmid itself.

    Conclusions:

    • IS-elements play a crucial role in the integrative recombination of the RP1 plasmid into the E. coli K12 chromosome.
    • The newly identified IS111 element, along with IS1-like and IS5-like elements, are involved in Hfr formation and chromosomal gene transfer.
    • RP1ts12 derivatives offer a tool for studying IS-element properties and detecting transposable elements in gram-negative bacteria.