Site‐specific insertion of gene cassettes into integrons

Christina M. Collis, Georgia Grammaticopoulos*, Jayne Briton, H. W. Stokes, Ruth M. Hall

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    173 Citations (Scopus)

    Abstract

    Site‐specific insertion of gene cassettes into the insert region of integrons has been demonstrated. Insertion was only observed if the integron DNA integrase was expressed in the recipient cell and if the cassette DNA was ligated prior to transformation. The essential ligation products were resistant to treatment with exonuclease III, indicating that they were closed circular molecules. Insertion of cassettes into integron fragments containing either no insert (one recombination site), or one gene cassette (two recombination sites), was demonstrated. In the latter case, insertion occurred predominantly at the core site located 5′ to the resident cassette, which corresponds to the only site available when no insert is present in the recipient. When DNA molecules including two gene cassettes were used, insertion of only one of the gene cassettes was generally observed, suggesting that resolution of the circular molecule to generate two independent circular cassettes occurred more rapidly than insertion into the recipient integron.

    Original languageEnglish
    Pages (from-to)41-52
    Number of pages12
    JournalMolecular Microbiology
    Volume9
    Issue number1
    DOIs
    Publication statusPublished - Jul 1993

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