Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis

Detalhes bibliográficos
Autor(a) principal: Croda, Julio Henrique Rosa
Data de Publicação: 2008
Outros Autores: Figueira, Cláudio Pereira, Wunder Junior, Elsio Augusto, Santos, Cleiton Silva, Reis, Mitermayer Galvão dos, Ko, Albert Icksang, Picardeau, Mathieu
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/1963
Resumo: Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
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spelling Croda, Julio Henrique RosaFigueira, Cláudio PereiraWunder Junior, Elsio AugustoSantos, Cleiton SilvaReis, Mitermayer Galvão dosKo, Albert IcksangPicardeau, Mathieu2011-04-25T19:41:54Z2011-04-25T19:41:54Z2008CRODA, J. et al. Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis. Infection and Immunity, v. 76, n.12, p.5826-5833, Dec. 2008.0019-9567https://www.arca.fiocruz.br/handle/icict/196310.1128/IAI.00989-08engTargeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilInstitut Pasteur. Unité de Biologie des Spirochètes. Paris, Françaof leptospirosis, remain largely unknown. This is mainly due to the lack of tools for genetically manipulating pathogenic Leptospira species. Thus, homologous recombination between introduced DNA and the corresponding chromosomal locus has never been demonstrated for this pathogen. Leptospiral immunoglobulin-like repeat (Lig) proteins were previously identified as putative Leptospira virulence factors. In this study, a ligB mutant was constructed by allelic exchange in L. interrogans; in this mutant a spectinomycin resistance (Spc(r)) gene replaced a portion of the ligB coding sequence. Gene disruption was confirmed by PCR, immunoblot analysis, and immunofluorescence studies. The ligB mutant did not show decrease virulence compared to the wild-type strain in the hamster model of leptospirosis. In addition, inoculation of rats with the ligB mutant induced persistent colonization of the kidneys. Finally, LigB was not required to mediate bacterial adherence to cultured cells. Taken together, our data provide the first evidence of site-directed homologous recombination in pathogenic Leptospira species. Furthermore, our data suggest that LigB does not play a major role in dissemination of the pathogen in the host and in the development of acute disease manifestations or persistent renal colonizationAntígenos de BactériasGenes BacterianosLeptospira interrogansLeptospiroseReação em Cadeia da PolimeraseMicroscopia de FluorescênciaCãesModelos Animais de Doençasinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZORIGINALTargeted Mutagenesis in Pathogenic Leptospira Species.pdfTargeted Mutagenesis in Pathogenic Leptospira Species.pdfapplication/pdf1112744https://www.arca.fiocruz.br/bitstream/icict/1963/1/Targeted%20Mutagenesis%20in%20Pathogenic%20Leptospira%20Species.pdff397f862db66eb259601c6984e3bb1e8MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81648https://www.arca.fiocruz.br/bitstream/icict/1963/2/license.txte095249ac7cacefbfe39684dfe45e706MD52TEXTTargeted Mutagenesis in Pathogenic Leptospira Species.pdf.txtTargeted Mutagenesis in Pathogenic Leptospira Species.pdf.txtExtracted texttext/plain47883https://www.arca.fiocruz.br/bitstream/icict/1963/5/Targeted%20Mutagenesis%20in%20Pathogenic%20Leptospira%20Species.pdf.txtcd1589ea0620b8deaaf647dfbebc6e54MD55THUMBNAILTargeted Mutagenesis in Pathogenic Leptospira Species.pdf.jpgTargeted Mutagenesis in Pathogenic Leptospira Species.pdf.jpgGenerated Thumbnailimage/jpeg2234https://www.arca.fiocruz.br/bitstream/icict/1963/4/Targeted%20Mutagenesis%20in%20Pathogenic%20Leptospira%20Species.pdf.jpgb9dc267f33d80fbbe7e51ee101166713MD54icict/19632023-03-15 14:32:59.618oai:www.arca.fiocruz.br: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Repositório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352023-03-15T17:32:59Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.pt_BR.fl_str_mv Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis
title Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis
spellingShingle Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis
Croda, Julio Henrique Rosa
Antígenos de Bactérias
Genes Bacterianos
Leptospira interrogans
Leptospirose
Reação em Cadeia da Polimerase
Microscopia de Fluorescência
Cães
Modelos Animais de Doenças
title_short Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis
title_full Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis
title_fullStr Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis
title_full_unstemmed Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis
title_sort Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis
author Croda, Julio Henrique Rosa
author_facet Croda, Julio Henrique Rosa
Figueira, Cláudio Pereira
Wunder Junior, Elsio Augusto
Santos, Cleiton Silva
Reis, Mitermayer Galvão dos
Ko, Albert Icksang
Picardeau, Mathieu
author_role author
author2 Figueira, Cláudio Pereira
Wunder Junior, Elsio Augusto
Santos, Cleiton Silva
Reis, Mitermayer Galvão dos
Ko, Albert Icksang
Picardeau, Mathieu
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Croda, Julio Henrique Rosa
Figueira, Cláudio Pereira
Wunder Junior, Elsio Augusto
Santos, Cleiton Silva
Reis, Mitermayer Galvão dos
Ko, Albert Icksang
Picardeau, Mathieu
dc.subject.decs.pt_BR.fl_str_mv Antígenos de Bactérias
Genes Bacterianos
Leptospira interrogans
Leptospirose
Reação em Cadeia da Polimerase
Microscopia de Fluorescência
Cães
Modelos Animais de Doenças
topic Antígenos de Bactérias
Genes Bacterianos
Leptospira interrogans
Leptospirose
Reação em Cadeia da Polimerase
Microscopia de Fluorescência
Cães
Modelos Animais de Doenças
description Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
publishDate 2008
dc.date.issued.fl_str_mv 2008
dc.date.accessioned.fl_str_mv 2011-04-25T19:41:54Z
dc.date.available.fl_str_mv 2011-04-25T19:41:54Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv CRODA, J. et al. Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis. Infection and Immunity, v. 76, n.12, p.5826-5833, Dec. 2008.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/1963
dc.identifier.issn.none.fl_str_mv 0019-9567
dc.identifier.doi.none.fl_str_mv 10.1128/IAI.00989-08
identifier_str_mv CRODA, J. et al. Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis. Infection and Immunity, v. 76, n.12, p.5826-5833, Dec. 2008.
0019-9567
10.1128/IAI.00989-08
url https://www.arca.fiocruz.br/handle/icict/1963
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