Tn951 derivatives designed for high-frequency plasmid-specific transposition and deletion mutagenesis

T Michiels, G Cornelis

    Research output: Contribution to journalArticlepeer-review

    Abstract

    We describe the construction of a system allowing high-frequency transposition and deletion mutagenesis with class-II transposons containing a kanamycin or a chloramphenicol-resistance marker. The system utilizes the transposition function of Tn3 and the resolution function of Tn951/Tn2501 which leads to an uncoupling of the resolution and repression functions. It consists of defective transposons inserted into conjugative, replication thermosensitive plasmids. The properties of the system are: easily selectable resistance markers, high transposition frequencies onto plasmids, low transposition frequencies onto the host chromosome, placement of the tnpA gene outside the transposons so that "second-generation" transposition does not occur, possibility to transpose the whole system onto other plasmid vectors with different selection strategies, consecutive use of two transposons for deletion mutagenesis and restriction mapping.
    Original languageEnglish
    Pages (from-to)175-81
    Number of pages7
    JournalGene
    Volume43
    Issue number3
    Publication statusPublished - 1986

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