In vivo transposition of mariner-based elements in enteric bacteria and mycobacteria
Department of Molecular Genetics and Microbiology
Medical Subject Headings
Bacteriophage lambda; Base Sequence; Conjugation, Genetic; Cyanobacteria; *DNA Transposable Elements; DNA-Binding Proteins; Escherichia coli; Mutagenesis, Insertional; Mycobacterium smegmatis; Mycobacterium tuberculosis; Open Reading Frames; Plasmids; Polymerase Chain Reaction; Promoter Regions, Genetic; Rhodobacter capsulatus; Streptomyces; Transposases
Microbiology | Molecular genetics
mariner family transposons are widespread among eukaryotic organisms. These transposons are apparently horizontally transmitted among diverse eukaryotes and can also transpose in vitro in the absence of added cofactors. Here we show that transposons derived from the mariner element Himar1 can efficiently transpose in bacteria in vivo. We have developed simple transposition systems by using minitransposons, made up of short inverted repeats flanking antibiotic resistance markers. These elements can efficiently transpose after expression of transposase from an appropriate bacterial promoter. We found that transposition of mariner-based elements in Escherichia coli produces diverse insertion mutations in either a targeted plasmid or a chromosomal gene. With Himar1-derived transposons we were able to isolate phage-resistant mutants of both E. coli and Mycobacterium smegmatis. mariner-based transposons will provide valuable tools for mutagenesis and genetic manipulation of bacteria that currently lack well developed genetic systems.
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Citation: Proc Natl Acad Sci U S A. 1999 Feb 16;96(4):1645-50.