IS Pst9, an ISL3-like insertion sequence from Pseudomonas stutzeri AN10 involved in catabolic gene inactivation Authors Joseph A. Christie-Oleza Microbiology, Department of Biology, University of the Balearic Islands, Palma, Mallorca Balbina Nogales Microbiology, Department of Biology, University of the Balearic Islands, Palma, Mallorca Celia Martín-Cardona Microbiology, Department of Biology, University of the Balearic Islands, Palma, Mallorca Mariana P. Lanfranconi Microbiology, Department of Biology, University of the Balearic Islands, Palma, Mallorca Sebastián Albertí Microbiology, Department of Biology, University of the Balearic Islands, Palma, Mallorca; Microbiology, Department of Biology, University of the Balearic Islands, Palma, Mallorca Jorge Lalucat Microbiology, Department of Biology, University of the Balearic Islands, Palma, Mallorca; Mediterranean Institute for Advanced Studies (IMEDEA), University of the Balearic Islands, Palma Rafael Bosch Microbiology, Department of Biology, University of the Balearic Islands, Palma, Mallorca Keywords: Pseudomonas stutzeri, insertion sequences, mobile elements, transposition, catabolic gene inactivation Abstract A novel insertion sequence (IS), ISPst9, from Pseudomonas stutzeri AN10 was cloned and characterized. ISPst9 is a typical bacterial IS, consisting of a 2472-bp element flanked by 24-bp perfect inverted repeats that generates 8-bp AT-rich target duplications upon insertion. The sequence also contains a gene that encodes an active transposase (TnpA) with significant amino acid identity to members of the ISL3 family. Southern blot analysis of digested genomic DNA of strain AN10 and its 4-chlorosalicylate-degrading derivative strain AN142 demonstrated that native ISPst9 transposes in multiple copies, with one of them responsible for the nahH insertional inactivation observed in strain AN142. Precise excision of ISPst9 yielded NahH+ revertants of AN142 at high frequencies (up to 10-6). In vivo transposition, mainly in multiple copies, of an ISPst9 derivative containing a KmR cassette cloned into a suicide vector was also demonstrated. Hybridization experiments carried out with different strains of P. stutzeri and with 292 phylogenetically distinct environmental isolates suggested that the presence of an ISPst9-like IS occurs in diverse bacteria together with the presence of aromatic hydrocarbon-degrading determinants. Downloads PDF Published 2009-02-25 Issue Vol. 11 No. 2 (2008) Section Research Articles License Submission of a manuscript to International Microbiology implies: that the work described has not been published before, including publication in the World Wide Web (except in the form of an Abstract or as part of a published lecture, review, or thesis); that it is not under consideration for publication elsewhere; that all the coauthors have agreed to its publication. The corresponding author signs for and accepts responsability for releasing this material and will act on behalf of any and all coauthors regarding the editorial review and publication process.If an article is accepted for publication in International Microbiology, the authors (or other copyright holder) must transfer to the journal the right–not exclusive–to reproduce and distribute the article including reprints, translations, photographic reproductions, microform, electronic form (offline, online) or any other reproductions of similar nature. Nevertheless, all article in International Microbiology will be available on the Internet to any reader at no cost. The journal allows users to freely download, copy, print, distribute, search, and link to the full text of any article, provided the authorship and source of the published article is cited. The copyright owner's consent does not include copying for new works, or resale. In these cases, the specific written permission of International Microbiology must first be obtained.Authors are requested to create a link to the published article on the journal's website. The link must be accompanied by the following text: "The original publication is available on LINK at <http://www.im.microbios.org>. Please use the appropiate URL for the article in LINK. Articles disseminated via LINK are indexed, abstracted, and referenced by many abstracting and information services, bibliographic networks, subscription agencies, library networks, and consortia.
