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Related Experiment Videos

Nucleotide sequence of an insertion element, IS1.

H Ohtsubo, E Ohtsubo

    Proceedings of the National Academy of Sciences of the United States of America
    |February 1, 1978
    PubMed
    Summary
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    Small plasmids revealed that the insertion sequence IS1 has 768 bases with inverted repeats at its ends. These findings suggest illegitimate recombination, potentially involving RNA polymerase, causes deletions in IS1.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • Plasmids like R100 are crucial genetic elements in bacteria.
    • Insertion sequences (IS) are mobile genetic elements that can cause mutations.
    • Understanding IS element dynamics is key to bacterial evolution.

    Purpose of the Study:

    • To characterize the structure of the insertion sequence IS1.
    • To investigate the mechanisms underlying deletions at the ends of IS1.
    • To explore the potential role of RNA polymerase in IS1 recombination.

    Main Methods:

    • Utilized deletion plasmids (PSM2, PSM1, PSM15) derived from R100.
    • Employed restriction enzyme digestion (EcoR1, HindIII, Hae III, Hpa II, Hha I, Hinf, AIu I).
    • Performed nucleotide sequencing to determine IS1 DNA sequence and structure.

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    Main Results:

    • IS1 was determined to be 768 base pairs long.
    • Inverted repeat sequences of approximately 30 bases were identified at both ends of IS1.
    • Deletions at the IS1 ends were attributed to illegitimate recombination.

    Conclusions:

    • The nucleotide sequence of IS1 and its flanking regions were elucidated.
    • Illegitimate recombination is the mechanism for IS1 end deletions.
    • Sequence analysis suggests a potential role for RNA polymerase in IS1 recombination hotspots.