Title

In vivo transposition of mariner-based elements in enteric bacteria and mycobacteria

UMMS Affiliation

Department of Molecular Genetics and Microbiology

Date

2-17-1999

Document Type

Article

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

Disciplines

Microbiology | Molecular genetics

Abstract

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.

Rights and Permissions

Citation: Proc Natl Acad Sci U S A. 1999 Feb 16;96(4):1645-50.

Related Resources

Link to Article in PubMed