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Journal of Bacteriology, December 1998, p. 6101-6106, Vol. 180, No. 23
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Structural and Functional Characterization of IS1358 from Vibrio cholerae

Sandrine Dumontier, Patrick Trieu-Cuot, and Patrick Berche*

INSERM U.411, Laboratoire de Microbiologie, Faculté de Médecine Necker- Enfants Malades, 75730 Paris Cedex 15, France

Received 15 May 1998/Accepted 29 September 1998

The new epidemic serovar O139 of Vibrio cholerae has emerged from the pandemic serovar O1 biotype El Tor through the replacement of a 22-kbp DNA region by a 40-kbp O139-specific DNA fragment. This O139-specific DNA fragment contains an insertion sequence that was described previously (U. H. Stroeher, K. E. Jedani, B. K. Dredge, R. Morona, M. H. Brown, L. E. Karageorgos, J. M. Albert, and P. A. Manning, Proc. Natl. Acad. Sci. USA 92:10374-10378, 1995) and designated IS1358O139. We studied the distribution of the IS1358 element in strains from various serovars by Southern analysis. Its presence was detected in strains from serovars O1, O2, O22, O139, and O155 but not in strains from serovars O15, O39, and O141. Furthermore, IS1358 was present in multiple copies in strains from serovars O2, O22, and O155. We cloned and sequenced four copies of IS1358 from V. cholerae O22 and one copy from V. cholerae O155. A comparison of their nucleotide sequences with those of O1 and O139 showed that they were almost identical. We constructed a transposon consisting of a kanamycin resistance gene flanked by two directly oriented copies of IS1358 to study the functionality of this element. Transposition of this element from a nonmobilizable plasmid onto the conjugative plasmid pOX38-Gen was detected in an Escherichia coli recA donor at a frequency of 1.2 × 10-8. Sequence analysis revealed that IS1358 duplicates 10 bp at its insertion site.

* Corresponding author. Mailing address: INSERM U.411, Laboratoire de Microbiologie, Faculté de Médecine Necker-Enfants Malades, 156 rue de Vaugirard, 75730 Paris Cedex 15, France. Phone: (33) 1 40 61 53 79. Fax: (33) 1 40 61 55 92. E-mail: berche{at}necker.fr.

Journal of Bacteriology, December 1998, p. 6101-6106, Vol. 180, No. 23
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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