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Team:Alberta/References/Publications/In vivo transposition of mariner based elements in enteric bacteria and mycobacteria
From 2009.igem.org
Eric J. Rubin, Brian J. Akerley, Veronica N. Novik, David J. Lampe, Robert N. Husson and John J. Mekalanos
PNAS February 16, 1999 vol. 96 no. 4 1645-1650
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.
Link: [http://www.pnas.org/content/96/4/1645.abstract PNAS]
