Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons.
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 found to be distributed around the plasmid, but insertions in certain regions occurred more frequently than in others.
D000667:Ampicillin / Q000494:pharmacology; D010583:Bacteriophage mu / Q000235:genetics*; D002872:Chromosome Deletion; D002876:Chromosomes, Bacterial; D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D007612:Kanamycin / Q000494:pharmacology; D007763:Lac Operon; D009154:Mutation; D010403:Penicillin Resistance; D010957:Plasmids; D014161:Transduction, Genetic
D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D007612:Kanamycin; D000667:Ampicillin