Recombinant PilS: cloning, expression and biochemical characterization of a Pil-Fimbriae subunit

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dc.contributorLab. Bacteriologiapt_BR
dc.contributorPrograma de Pós-Graduação em Ciências – Toxinologia (PPGTox)pt_BR
dc.contributorLab. de Toxinologia Aplicadapt_BR
dc.contributorLaboratório Estratégico de Diagnóstico Molecular (LEDM)pt_BR
dc.contributor.authorMunhoz, Danielle Diaspt_BR
dc.contributor.authorSilva, Jessika Cristina Alves dapt_BR
dc.contributor.authorFreitas, Nataliapt_BR
dc.contributor.authorIwai, Leo Keipt_BR
dc.contributor.authorAires, Karina A.pt_BR
dc.contributor.authorOzaki, Christiane Y.pt_BR
dc.contributor.authorSouza, Cristiane S.pt_BR
dc.contributor.authorRocha, Letícia Barbozapt_BR
dc.contributor.authorSilva, Míriam Aparecida dapt_BR
dc.contributor.authorHenrique, Izabella de Macedopt_BR
dc.contributor.authorElias, Waldir Pereirapt_BR
dc.contributor.authorCarvalho, Eneaspt_BR
dc.contributor.authorMorganti, Ligiapt_BR
dc.contributor.authorChura-Chambi, Rosa Mariapt_BR
dc.contributor.authorPiazza, Roxane Maria Fontespt_BR
dc.date.accessioned2022-06-23T13:03:44Z-
dc.date.available2022-06-23T13:03:44Z-
dc.date.issued2022pt_BR
dc.identifier.citationMunhoz DD, Silva JCA, Freitas N, Iwai LK, Aires KA., Ozaki CY., et al. Recombinant PilS: cloning, expression and biochemical characterization of a Pil-Fimbriae subunit. Microorganisms. 2022 June;10(6):1174. doi:10.3390/microorganisms10061174.pt_BR
dc.identifier.urihttps://repositorio.butantan.gov.br/handle/butantan/4402-
dc.description.abstractPil-fimbriae is a type IV pili member, which is a remarkably versatile component with a wide variety of functions, including motility, attachment to different surfaces, electrical conductance, DNA acquisition, and secretion of a broad range of structurally distinct protein substrates. Despite the previous functional characterization of Pil, more studies are required to understand the regulation of Pil expression and production, since the exact mechanisms involved in these steps are still unknown. Therefore it is extremely important to have a protein with the correct secondary and tertiary structure that will enable an accurate characterization and a specific antisera generation. For this reason, the aim of this work was to generate potential tools for further investigations to comprehend the mechanisms involved in Pil regulation and its role in pathogenic E. coli infections with the obtaining of a precise native-like recombinant PilS and the corresponding antisera. The pilS gene was successfully cloned into an expression vector, and recombinant PilS (rPilS) was efficiently solubilized and purified by metal affinity chromatography. Protein characterization analyses indicated that rPilS presented native-like secondary and tertiary structures after the refolding process. The generated anti-rPilS sera efficiently recognized recombinant and native proteins from atypical enteropathogenic E. coli strains.pt_BR
dc.description.sponsorship(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulopt_BR
dc.description.sponsorship(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológicopt_BR
dc.description.sponsorship(CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorpt_BR
dc.format.extent1174pt_BR
dc.language.isoEnglishpt_BR
dc.relation.ispartofMicroorganismspt_BR
dc.rightsOpen accesspt_BR
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/pt_BR
dc.titleRecombinant PilS: cloning, expression and biochemical characterization of a Pil-Fimbriae subunitpt_BR
dc.typeArticlept_BR
dc.rights.licenseCC BYpt_BR
dc.identifier.doi10.3390/microorganisms10061174pt_BR
dc.identifier.urlhttps://doi.org/10.3390/microorganisms10061174pt_BR
dc.contributor.external(IPEN) Instituto de Pesquisas Energéticas e Nuclearespt_BR
dc.contributor.externalHospital Israelita Albert Einsteinpt_BR
dc.contributor.external(USP) Universidade de São Paulopt_BR
dc.identifier.citationvolume10pt_BR
dc.identifier.citationissue6pt_BR
dc.subject.keywordadhesinpt_BR
dc.subject.keywordPil fimbriaept_BR
dc.subject.keywordrecombinant proteinpt_BR
dc.subject.keywordcloningpt_BR
dc.subject.keywordexpressionpt_BR
dc.subject.keywordbiochemical characterizationpt_BR
dc.relation.ispartofabbreviatedMicroorganismspt_BR
dc.identifier.citationabntv. 10, n. 6, 1174, jun. 2022pt_BR
dc.identifier.citationvancouver2022 June;10(6):1174pt_BR
dc.contributor.butantanMunhoz, Danielle Dias|:Desvinculado|:Lab. Bacteriologia|:PrimeiroAutorpt_BR
dc.contributor.butantanSilva, Jessika Cristina Alves da|:Aluno|:Lab. Bacteriologia:Programa de Pós-Graduação em Ciências – Toxinologia (PPGTox)pt_BR
dc.contributor.butantanFreitas, Natalia|:Desvinculado|:Lab. Bacteriologiapt_BR
dc.contributor.butantanIwai, Leo Kei|:Pesquisador:Docente Colaborador PPGTox|:Lab. de Toxinologia Aplicadapt_BR
dc.contributor.butantanAires, Karina A.|:Desvinculado|:Lab. Bacteriologiapt_BR
dc.contributor.butantanKarina A. Aires|:Desvinculado|:Lab. Bacteriologiapt_BR
dc.contributor.butantanSouza, Cristiane S.|:Desvinculado|:Lab. Bacteriologiapt_BR
dc.contributor.butantanRocha, Letícia Barboza|:Outros|:Laboratório Estratégico de Diagnóstico Molecular (LEDM)pt_BR
dc.contributor.butantanSilva, Míriam Aparecida da|:Técnico|:Lab. Bacteriologiapt_BR
dc.contributor.butantanHenrique, Izabella de Macedo|:Desvinculado|:Lab. Bacteriologiapt_BR
dc.contributor.butantanElias, Waldir Pereira|:Pesquisador|:Lab. Bacteriologiapt_BR
dc.contributor.butantanCarvalho, Eneas|:Pesquisador|:Lab. Bacteriologiapt_BR
dc.contributor.butantanChura-Chambi, Rosa Maria|:Outros|:Lab. Bacteriologiapt_BR
dc.contributor.butantanPiazza, Roxane Maria Fontes|:Pesquisador:Docente Permanente PPGTox|:Lab. Bacteriologiapt_BR
dc.sponsorship.butantan(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo¦¦2004/12136-5pt_BR
dc.sponsorship.butantan(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo¦¦2009/14845-7pt_BR
dc.sponsorship.butantan(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológico¦¦470648/2008-2pt_BR
dc.sponsorship.butantan(CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior¦¦pt_BR
dc.identifier.bvsccBR78.1pt_BR
dc.identifier.bvsdbIBProdpt_BR
dc.description.dbindexedYespt_BR
item.openairetypeArticle-
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item.languageiso639-1English-
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