title: Efficient generation of unmarked deletions in Legionella pneumophila.
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.
authors.affiliation: Department 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.
authors.forename: Andrew
authors.initials: A
authors.lastname: Bryan
authors.forename: Kaoru
authors.initials: K
authors.lastname: Harada
authors.forename: Michele S
authors.initials: MS
authors.lastname: Swanson
mesh_terms: D004926:Escherichia coli / Q000235:genetics; D005827:Genetics, Microbial / Q000379:methods*; D016952:Legionella pneumophila / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000379:methods; D010957:Plasmids; D011995:Recombination, Genetic; D012641:Selection, Genetic; D017384:Sequence Deletion; D014169:Transformation, Bacterial