Genetic tools for studying Capnocytophaga canimorsus.

Applied and Environmental Microbiology
Manuela Mally, Guy R Cornelis

Abstract

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 thes...Continue Reading

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Citations

Mar 17, 2009·Microbes and Infection·Manuela MallyGuy R Cornelis
Jul 25, 2015·Anaerobe·Zohreh Tamanai-ShacooriAnne Jolivet-Gougeon
Aug 10, 2010·Journal of Microbiological Methods·M Paula Goetting-Minesky, J Christopher Fenno
Nov 5, 2014·Infection and Immunity·Pablo ManfrediGuy R Cornelis
Dec 16, 2016·Scientific Reports·Francesco RenziGuy R Cornelis
Dec 9, 2015·Infection and Immunity·Francesco RenziSimon J Ittig
Apr 7, 2017·Journal of Clinical Microbiology·Estelle HessGuy R Cornelis
Nov 1, 2016·MBio·Frédéric LauberFrancesco Renzi
Aug 30, 2017·Applied Microbiology and Biotechnology·Yongtao Zhu, Mark J McBride
Jun 29, 2018·Applied Microbiology and Biotechnology·MaFeng LiuAnChun Cheng
Dec 29, 2016·Journal of Thrombosis and Haemostasis : JTH·K HackG R Cornelis
Mar 10, 2009·Veterinary Microbiology·Wim Gaastra, Len J A Lipman

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