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Structural requirement for IS50-mediated gene transposition.

D E Berg

    Proceedings of the National Academy of Sciences of the United States of America
    |February 1, 1983
    PubMed
    Summary
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    Transposon IS50, part of Tn5, likely moves DNA conservatively, not by forming cointegrates. This study examined transposition products to understand the mechanism of DNA movement.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • Transposable elements mediate DNA rearrangement.
    • Replicative transposition typically forms cointegrates, joining donor and target DNA.
    • IS50 elements are terminal repeats of Tn5, mediating transposition.

    Purpose of the Study:

    • To investigate whether IS50 transposition generates cointegrates.
    • To determine the mechanism of DNA movement mediated by IS50.

    Main Methods:

    • Transposition experiments using pBR322::Tn5 plasmids and an F factor in recA-Escherichia coli.
    • Analysis of transposition products using restriction endonuclease digestion.

    Main Results:

    • Transposition of kanamycin resistance (Kanr) was often not accompanied by ampicillin resistance (Ampr) movement from monomeric plasmids.

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  • From dimeric plasmids, half of Kanr transpositions included Ampr movement.
  • No true cointegrates were observed; analyzed chimeras contained single copies of vector DNA bracketed by IS50/Tn5 elements.

    • Conclusions:

    • IS50 transposition does not appear to generate cointegrates.
    • Evidence suggests IS50 transposition proceeds via a conservative mechanism, moving only a segment of the donor replicon.