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Applied and Environmental Microbiology, January 2008, p. 319-322, Vol. 74, No. 1
0099-2240/08/$08.00+0     doi:10.1128/AEM.02172-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Conjugative Transfer between Escherichia coli and Leptospira spp. as a New Genetic Tool{triangledown}

Mathieu Picardeau*

Unité de Biologie des Spirochètes, Institut Pasteur, Paris, France

Received 24 September 2007/ Accepted 30 October 2007

Our understanding of leptospiral pathogenesis, which remains poorly understood, depends on reliable genetic tools for functional analysis of genes in pathogenic strains. In this study, we report the first demonstration of conjugation between Escherichia coli and Leptospira spp. by using RP4 derivative conjugative plasmids. The DNA transfer described here was due to authentic conjugation, as shown by the requirement for cell-to-cell contact and the resistance of DNA transfers to the addition of DNase I. Transposition via conjugation of a plasmid delivering Himar1 yielded frequencies ranging from 1 x 10–6 to 8.5 x 10–8 transconjugants/recipient cell in the saprophyte L. biflexa and the pathogen L. interrogans, respectively. Analysis of mutants indicated that transposition occurs randomly, and at single sites in the genome of these strains, allowing the utilization of this system to generate libraries of transposon mutants.

* Mailing address: Unité de Biologie des Spirochètes, Institut Pasteur, 28 rue du docteur Roux, 75724 Paris Cedex 15, France. Phone: 33 (1) 45 68 83 68. Fax: 33 (1) 40 61 30 01. E-mail: mpicard{at}pasteur.fr

{triangledown} Published ahead of print on 9 November 2007.

Applied and Environmental Microbiology, January 2008, p. 319-322, Vol. 74, No. 1
0099-2240/08/$08.00+0     doi:10.1128/AEM.02172-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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