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Journal of Bacteriology, February 2003, p. 918-928, Vol. 185, No. 3
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.3.918-928.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Recombination Activity of a Distinctive Integron-Gene Cassette System Associated with Pseudomonas stutzeri Populations in Soil

Andrew J. Holmes,1* Marita P. Holley,1 Andrew Mahon,2 Blair Nield,2 Michael Gillings,1 and H. W. Stokes2

Key Centre for Biodiversity and Bioresources,1 Department of Biological Sciences, Macquarie University, Sydney NSW 2109, Australia2

Received 15 July 2002/ Accepted 9 November 2002

Class 1 integrons have strongly influenced the evolution of multiple antibiotic resistance. Diverse integrons have recently been detected directly in a range of natural environments. In order to characterize the properties of these environmental integrons, we sought to isolate organisms containing integrons from soils, which resulted in the isolation of Pseudomonas stutzeri strain Q. Further isolation efforts targeted at this species resulted in recovery of two other strains (P and BAM). 16S rRNA sequences and chromosome mapping showed that these three strains are very closely related clonal variants in a single genomovar of P. stutzeri. Only strains Q and BAM were found to contain an integron and an associated gene cassette array. The intI and attI components of these strains showed 99 and 90% identity, respectively. The structure of these integrons and their associated gene cassettes was similar to that reported previously for other integron classes. The two integrons contained nonoverlapping sets of cassette-associated genes. In contrast, many of the cassette-associated recombination sites in the two integrons were similar and were considered to constitute a distinct subfamily consisting of 59-base element (59-be) recombination sites (the Pseudomonas subfamily). The recombination activity of P. stutzeri integron components was tested in cointegrate assays. IntIPstQ was shown to catalyze site-specific recombination between its cognate attI site and 59-be sites from antibiotic resistance gene cassettes. While IntIPstQ did not efficiently mediate recombination between members of the Pseudomonas 59-be subfamily and other 59-be types, the former sites were functional when they were tested with IntI1. We concluded that integrons present in P. stutzeri possess recombination activity and represent a hot spot for genomic diversity in this species.

* Corresponding author. Present address: School of Molecular and Microbial Biosciences, The University of Sydney, Sydney, NSW 2006, Australia. Phone: (612) 9351 2530. Fax: (612) 9351 4571. E-mail: A.Holmes{at}mmb.usyd.edu.au.

Journal of Bacteriology, February 2003, p. 918-928, Vol. 185, No. 3
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.3.918-928.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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