Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons

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Abstract
Small bacteriophage Mu transposable elements containing the lac operon structural genes were constructed to facilitate the isolation and use of Mu insertions, and lac gene fusions. These mini-Mu elements have selectable genes for either ampicillin or kanamycin resistance, and can be used to form both transcriptional and translational lac gene fusions. Some of the mini-Mu-lac elements constructed are deleted for the Mu A and B transposition genes, and form stable insertions that cannot undergo transposition unless complemented for these functions. A procedure was developed for selecting mini-Mu insertions specifically into plasmids, including commonly used high-copy-number cloning vectors such as pBR322. Mu insertions in pBR322 were distributed around the plasmid, but insertions in certain regions occurred more frequently than in others.