Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Outros Autores: | , , , , , |
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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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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eng |
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eng |
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info:eu-repo/semantics/openAccess |
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openAccess |
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