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Applied and Environmental Microbiology, October 2008, p. 6369-6377, Vol. 74, No. 20
0099-2240/08/$08.00+0     doi:10.1128/AEM.01218-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Genetic Tools for Studying Capnocytophaga canimorsus{triangledown}

Manuela Mally and Guy R. Cornelis*

Infection Biology, Biozentrum, University of Basel, Klingelbergstrasse 50/70, CH-4056 Basel, Switzerland

Received 2 June 2008/ Accepted 13 August 2008

Capnocytophaga canimorsus, a commensal bacterium from canine oral flora, has been isolated throughout the world from severe human infections caused by dog bites. Due to the low level of evolutionary relationship to Proteobacteria, genetic methods suitable for the genus Capnocytophaga needed to be established. Here, we show that Tn4351, derived from Bacteroides fragilis, could be introduced by conjugation into C. canimorsus and conferred resistance to erythromycin. By mapping and sequencing a naturally occurring plasmid isolated from a clinical isolate of C. canimorsus, we identified a repA gene that allowed us to construct Escherichia coli-Capnocytophaga shuttle vectors. Most commonly used antibiotic markers were not functional in C. canimorsus, but cefoxitin (cfxA), tetracycline (tetQ), and erythromycin (ermF) resistances could be used as markers for plasmid maintenance in C. canimorsus and even in some other Capnocytophaga spp. Shuttle vectors were introduced into C. canimorsus either by conjugation using the origin of transfer (oriT) of RP4 or by electrotransformation. Taking advantage of the promoter of ermF, an expression vector was constructed. Finally, a method that allows site-directed mutagenesis is described. All these genetic tools pave the way, not only for molecular studies of the pathogenesis of C. canimorsus, but also for studies of other oral Capnocytophaga species.

* Corresponding author. Mailing address: Biozentrum, Klingelbergstrasse 50-70, CH-4056 Basel, Switzerland. Phone: 41 61 267 21 10. Fax: 41 61 267 21 18. E-mail: guy.cornelis{at}unibas.ch

{triangledown} Published ahead of print on 22 August 2008.

Applied and Environmental Microbiology, October 2008, p. 6369-6377, Vol. 74, No. 20
0099-2240/08/$08.00+0     doi:10.1128/AEM.01218-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Shin, H., Mally, M., Meyer, S., Fiechter, C., Paroz, C., Zaehringer, U., Cornelis, G. R. (2009). Resistance of Capnocytophaga canimorsus to Killing by Human Complement and Polymorphonuclear Leukocytes. Infect. Immun. 77: 2262-2271 [Abstract] [Full Text]  


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