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Applied and Environmental Microbiology, April 2005, p. 1822-1828, Vol. 71, No. 4
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.4.1822-1828.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

High-Temperature-Induced Transposition of Insertion Elements in Burkholderia multivorans ATCC 17616

Yoshiyuki Ohtsubo,* Hiroyuki Genka, Harunobu Komatsu, Yuji Nagata, and Masataka Tsuda

Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, Japan

Received 29 July 2004/ Accepted 5 November 2004

An efficient and quantitative method to analyze the transposition of various insertion sequence (IS) elements in Burkholderia multivorans ATCC 17616 was devised. pGEN500, a plasmid carrying a Bacillus subtilis-derived sacB gene, was introduced into ATCC 17616 cells, and 25% of their sucrose-resistant derivatives were found to carry various IS elements on pGEN500. A PCR-based experimental protocol, in which a mixture of several specific primer pairs was used, revealed that pGEN500 captured, in addition to five previously reported IS elements (IS401, IS402, IS406, IS407, and IS408), three novel IS elements, ISBmu1, ISBmu2, and ISBmu3. The global transposition frequency of these IS elements was enhanced more than sevenfold under a high-temperature condition (42°C) but not under oxidative stress or starvation conditions. To our knowledge, this is the first report demonstrating the elevated transposition activities of several IS elements at a high temperature. The efficient experimental protocol developed in this study will be useful in quantitatively and simultaneously investigating various IS elements, as well as in capturing novel functional mobile elements from a wide variety of bacteria.

* Corresponding author. Mailing address: Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Sendai 980-8577, Japan. Phone: 81-22-217-5743. Fax: 81-22-217-5699. E-mail: yohtsubo{at}ige.tohoku.ac.jp.

Applied and Environmental Microbiology, April 2005, p. 1822-1828, Vol. 71, No. 4
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.4.1822-1828.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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