Efficient generation of unmarked deletions in Legionella pneumophila
Andrew Bryan1, Kaoru Harada, Michele S Swanson
1Department of Microbiology and Immunology, University of Michigan Medical School, 5641 Medical Sciences Building II, 1150 West Medical Center Dr., Ann Arbor, MI 48109-5620, USA.
Abstract:
Unmarked gene deletions facilitate studies of Legionella pneumophila multicomponent processes, such as motility and exonuclease activity. For this purpose, FRT-flanked alleles constructed in Escherichia coli using λ-Red recombinase were transferred to L. pneumophila by natural transformation. Resistance cassettes were then efficiently excised using the Flp site-specific recombinase encoded on a plasmid that is readily lost.
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