Development of transcriptional fusions to assess Leptospira interrogans promoter activity

Cerqueira, Gustavo M., Souza, Natalie M., Araújo, Eduardo R., Barros, Aline T., Morais, Zenaide M., Vasconcellos, Sílvio A., and Nascimento, Ana L.T.O. (2011) Development of transcriptional fusions to assess Leptospira interrogans promoter activity. PLoS ONE, 6. pp. 1-16.

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Abstract

Background: Leptospirosis is a zoonotic infectious disease that affects both humans and animals. The existing genetic tools for Leptospira spp. have improved our understanding of the biology of this spirochete as well as the interaction of pathogenic leptospires with the mammalian host. However, new tools are necessary to provide novel and useful information to the field.

Methodology and Principal Findings: A series of promoter-probe vectors carrying a reporter gene encoding green fluorescent protein (GFP) were constructed for use in L. biflexa. They were tested by constructing transcriptional fusions between the lipL41, Leptospiral Immunoglobulin-like A (ligA) and Sphingomyelinase 2 (sph2) promoters from L. interrogans and the reporter gene. ligA and sph2 promoters were the most active, in comparison to the lipL41 promoter and the non-induced controls. The results obtained are in agreement with LigA expression from the L. interrogans Fiocruz L1-130 strain.

Conclusions: The novel vectors facilitated the in vitro evaluation of L. interrogans promoter activity under defined growth conditions which simulate the mammalian host environment. The fluorescence and rt-PCR data obtained closely reflected transcriptional regulation of the promoters, thus demonstrating the suitability of these vectors for assessing promoter activity in L. biflexa.

Item ID: 37414
Item Type: Article (Research - C1)
ISSN: 1932-6203
Additional Information:

© 2011 Cerqueira et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Date Deposited: 25 Feb 2015 03:21
FoR Codes: 06 BIOLOGICAL SCIENCES > 0605 Microbiology > 060502 Infectious Agents @ 50%
06 BIOLOGICAL SCIENCES > 0605 Microbiology > 060503 Microbial Genetics @ 50%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100%
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