Expanding the Genetic Toolbox for Leptospira Species by Generation of Fluorescent Bacteria - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Applied and Environmental Microbiology Année : 2010

Expanding the Genetic Toolbox for Leptospira Species by Generation of Fluorescent Bacteria

Résumé

Our knowledge of the genetics and molecular basis of the pathogenesis associated with Leptospira , in comparison to those of other bacterial species, is very limited. An improved understanding of pathogenic mechanisms requires reliable genetic tools for functional genetic analysis. Here, we report the expression of gfp and mRFP1 genes under the control of constitutive spirochetal promoters in both saprophytic and pathogenic Leptospira strains. We were able to reliably measure the fluorescence of Leptospira by fluorescence microscopy and a fluorometric microplate reader-based assay. We showed that the expression of the gfp gene had no significant effects on growth in vivo and pathogenicity in L. interrogans . We constructed an expression vector for L. biflexa that contains the lacI repressor, an inducible lac promoter, and gfp as the reporter, demonstrating that the lac system is functional in Leptospira . Green fluorescent protein (GFP) expression was induced by the addition of isopropyl-β- d -thiogalactopyranoside (IPTG) in L. biflexa transformants harboring the expression vector. Finally, we showed that GFP can be used as a reporter to assess promoter activity in different environmental conditions. These results may facilitate further advances for studying the genetics of Leptospira spp.

Dates et versions

hal-03324588 , version 1 (23-08-2021)

Identifiants

Citer

Florence Aviat, Leyla Slamti, Gustavo M. Cerqueira, Kristel Lourdault, Mathieu Picardeau. Expanding the Genetic Toolbox for Leptospira Species by Generation of Fluorescent Bacteria. Applied and Environmental Microbiology, 2010, 76 (24), pp.8135-8142. ⟨10.1128/AEM.02199-10⟩. ⟨hal-03324588⟩

Collections

PASTEUR ANR
11 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More