title: Site-specific insertion of IS492 in Pseudoalteromonas atlantica.
abstract: Reversible insertion of IS492 at a site within epsG on the Pseudoalteromonas atlantica chromosome controls peripheral extracellular polysaccharide production and biofilm formation by P. atlantica. High-frequency precise excision of IS492 from epsG requires 5 and 7 bp of flanking DNA, suggesting that IS492 transposition involves a site-specific recombination mechanism. The site specificity of IS492 insertion was examined in P. atlantica and shown to be specific for a 7-bp target, 5'-CTTGTTA-3'. Characterization of numerous insertion events at the target site in epsG indicated that insertion is also orientation specific. The frequency of IS492 insertion at the epsG target site (2.7 x 10(-7)/cell/generation), determined by quantitative PCR, is 4 to 5 orders of magnitude lower than the frequency of IS492 precise excision from the same site. Comparison of insertion sites for IS492 and the highly related ISPtu2 from Pseudoalteromonas tunicata suggests DNA sequence and/or structural features that may contribute to site recognition and recombination by the transposase of IS492.
authors.affiliation: Department of Microbiology, University of Georgia, Athens, Georgia 30602, USA.
authors.forename: Brian P
authors.initials: BP
authors.lastname: Higgins
authors.forename: Adam C
authors.initials: AC
authors.lastname: Popkowski
authors.forename: Peter R
authors.initials: PR
authors.lastname: Caruana
authors.forename: Anna C
authors.initials: AC
authors.lastname: Karls
mesh_terms: D001483:Base Sequence; D002876:Chromosomes, Bacterial; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D031161:Pseudoalteromonas / Q000235:genetics* / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial
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