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Journal of Bacteriology, October 2009, p. 6408-6414, Vol. 191, No. 20
0021-9193/09/$08.00+0     doi:10.1128/JB.00771-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Site-Specific Insertion of IS492 in Pseudoalteromonas atlantica{triangledown}

Brian P. Higgins,{dagger} Adam C. Popkowski,{ddagger} Peter R. Caruana,§ and Anna C. Karls*

Department of Microbiology, University of Georgia, Athens, Georgia

Received 13 June 2009/ Accepted 3 August 2009

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.

* Corresponding author. Mailing address: 1000 Cedar Street, 527 Biological Sciences Bldg., Athens, GA 30602. Phone: (706) 583-0822. Fax: (706) 542-2674. E-mail: akarls{at}uga.edu

{triangledown} Published ahead of print on 14 August 2009.

{dagger} Present address: Department of Ecology and Evolutionary Biology, Princeton University, Princeton, NJ.

{ddagger} Present address: Mercer University School of Medicine, Macon, GA.

§ Present address: University of Virginia School of Medicine, Charlottesville, VA.

Journal of Bacteriology, October 2009, p. 6408-6414, Vol. 191, No. 20
0021-9193/09/$08.00+0     doi:10.1128/JB.00771-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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