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Related Experiment Videos

Plasmids containing insertion elements are potential transposons.

E Ohtsubo, M Zenilman, H Ohtsubo

    Proceedings of the National Academy of Sciences of the United States of America
    |February 1, 1980
    PubMed
    Summary
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    This study shows how insertion sequence IS1 on a plasmid can integrate into another plasmid, forming recombinant DNA. This process, independent of recA, suggests plasmids with translocatable elements can act as transposons.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • Plasmids are extrachromosomal DNA elements crucial in bacterial genetics.
    • Insertion sequences (IS) are mobile genetic elements that can mediate DNA rearrangements.
    • Understanding plasmid recombination is key to deciphering gene transfer and evolution.

    Purpose of the Study:

    • To investigate the in vivo recombination mechanism between two plasmids, pHS1 and pSM1.
    • To characterize the role of the insertion element IS1 in plasmid integration.
    • To determine if this recombination process is dependent on the recA gene.

    Main Methods:

    • Utilized temperature-sensitive replication mutant plasmid pHS1 and small plasmid pSM1 containing IS1.
    • Selected for tetracycline resistance at 42°C to identify recombinant plasmids.

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  • Employed electron microscopy and restriction endonuclease digestion for physical structure analysis.
  • Conducted nucleotide sequence analysis of recombinant and parental DNA.

    • Main Results:

    • Identified recombinant plasmids formed through IS1 integration into pHS1, independent of recA function.
    • Observed IS1 integration at multiple sites on pHS1, resulting in duplicated IS1 sequences at junctions.
    • Confirmed target site duplication (nine nucleotides) at each IS1 junction via sequence analysis.

    Conclusions:

    • Plasmid recombination mediated by IS1 involves target DNA duplication, mimicking transposition.
    • Plasmids harboring translocatable DNA elements, like IS1, can function as transposons.
    • The recA-independent nature of this recombination highlights alternative pathways for genetic exchange.