title: Development of transcriptional fusions to assess Leptospira interrogans promoter activity.
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.
  
  
  RESULTS
  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.
authors: Centro de Biotecnologia, Instituto Butantan, São Paulo, Brazil. cerqueiragm@yahoo.com.br
authors: Gustavo M
authors: GM
authors: Cerqueira
authors: Natalie M
authors: NM
authors: Souza
authors: Eduardo R
authors: ER
authors: Araújo
authors: Aline T
authors: AT
authors: Barros
authors: Zenaide M
authors: ZM
authors: Morais
authors: Sílvio A
authors: SA
authors: Vasconcellos
authors: Ana L T O
authors: AL
authors: Nascimento
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D050939:Gene Fusion; D005798:Genes, Bacterial; D007921:Leptospira interrogans / Q000235:genetics*; D008856:Microscopy, Fluorescence; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
chemical_list: D017931:DNA Primers