Expanding the genetic toolbox for Leptospira species by generation of fluorescent bacteria

Aviat, Florence, Slamti, Leyla, Cerqueira, Gustavo M., Lourdault, Kristel, and Picardeau, Mathieu (2010) Expanding the genetic toolbox for Leptospira species by generation of fluorescent bacteria. Applied and Environmental Microbiology, 76 (24). pp. 8135-8142.

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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.

Item ID: 37419
Item Type: Article (Research - C1)
ISSN: 1098-5336
Funders: CAPES Foundation, Brazilian Ministry of Education, Institut Pasteur, French Ministry of Research (FMR), Fiocruz-Pasteur Scientific Cooperation Agreement
Projects and Grants: FMR ANR-08-MIE-018
Date Deposited: 12 Mar 2015 03:51
FoR Codes: 06 BIOLOGICAL SCIENCES > 0605 Microbiology > 060503 Microbial Genetics @ 50%
06 BIOLOGICAL SCIENCES > 0605 Microbiology > 060502 Infectious Agents @ 50%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100%
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