Compostos indutores e genes regulando a expressão da bomba de efluxo Tap em Mycobacterium bovis BCG
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFPel - Guaiaca |
Texto Completo: | http://guaiaca.ufpel.edu.br/handle/123456789/1222 |
Resumo: | Transport proteins related to drug efflux play an important role not only in the acquisition of drug-resistant phenotypes, but also in virulence of M. tuberculosis. The main objective of this study was to analyze the regulation of genes encoding the protein Rv1258c Tap considering the expression of genes Rv1255c and Rv1257c. RNA was extracted from cultures of M. bovis BCG overexpressing Rv1255c and Rv1257c, as well as a control strain containing the vector pVV16, and qRT-PCR of the Rv1258c gene was performed. RT-PCR was performed using RNA isolated previously from M. tuberculosis CDC1551, in order to verify that Rv1255c, Rv1256c, Rv1257c and Rv1258c genes are all connected to a transcript. The strain M. bovis BCG, expressing a red fluorescent protein under control of the promoter Rv1258c (pVVTapPro) was grown in the presence of streptomycin and glycine to verify the induction of the promoter. The qRT-PCR showed a downregulation of the gene in the strain overexpressing Rv1258c and Rv1257c, but no difference was observed in the strain overexpressing Rv1255c compared with the control. An increase in fluorescence was observed in the strain containing the promoter of the tap in the presence of 1 mg/L of streptomycin in comparison with the control. A two-fold induction of gene Tap was also observed in the presence of 1.6 mM glycine. The results suggest a role for Rv1257c in the regulation Tap expression. Moreover, the induction of Tap by streptomycin supports the importance of this protein in multidrugresistant M. tuberculosis. The effect of glycine on Tap promoter suggest a physiological role in cellular detoxification for this efflux pump. In conclusion, this study reinforces the need to obtain a deeper knowledge about the Tap protein operon and its regulation, in order to assist in the development of inhibitors of this efflux pump and possibly other mechanisms of induced resistance. |
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2014-08-20T13:32:48Z2014-01-242014-08-20T13:32:48Z2013-08-28FELIX, Carolina Rodrigues. Inductors and genes that regulate expression of the Tap efflux pump in Mycobacterium bovis BCG. 2013. 52 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2013.http://guaiaca.ufpel.edu.br/handle/123456789/1222Transport proteins related to drug efflux play an important role not only in the acquisition of drug-resistant phenotypes, but also in virulence of M. tuberculosis. The main objective of this study was to analyze the regulation of genes encoding the protein Rv1258c Tap considering the expression of genes Rv1255c and Rv1257c. RNA was extracted from cultures of M. bovis BCG overexpressing Rv1255c and Rv1257c, as well as a control strain containing the vector pVV16, and qRT-PCR of the Rv1258c gene was performed. RT-PCR was performed using RNA isolated previously from M. tuberculosis CDC1551, in order to verify that Rv1255c, Rv1256c, Rv1257c and Rv1258c genes are all connected to a transcript. The strain M. bovis BCG, expressing a red fluorescent protein under control of the promoter Rv1258c (pVVTapPro) was grown in the presence of streptomycin and glycine to verify the induction of the promoter. The qRT-PCR showed a downregulation of the gene in the strain overexpressing Rv1258c and Rv1257c, but no difference was observed in the strain overexpressing Rv1255c compared with the control. An increase in fluorescence was observed in the strain containing the promoter of the tap in the presence of 1 mg/L of streptomycin in comparison with the control. A two-fold induction of gene Tap was also observed in the presence of 1.6 mM glycine. The results suggest a role for Rv1257c in the regulation Tap expression. Moreover, the induction of Tap by streptomycin supports the importance of this protein in multidrugresistant M. tuberculosis. The effect of glycine on Tap promoter suggest a physiological role in cellular detoxification for this efflux pump. In conclusion, this study reinforces the need to obtain a deeper knowledge about the Tap protein operon and its regulation, in order to assist in the development of inhibitors of this efflux pump and possibly other mechanisms of induced resistance.Proteínas transportadoras relacionada ao efluxo de drogas desempenham um papel importante não só na aquisição de fenótipos resistentes aos fármacos, mas também na virulência de M. tuberculosis. O principal objetivo deste estudo foi analisar a regulação do gene Rv1258c codificador da proteína Tap considerando a expressão dos genes Rv1255c e Rv1257c. RNA foi extraído a partir de culturas de M. bovis BCG superexpressando Rv1255c e Rv1257c, bem como de uma cepa controle contendo o vector pVV16 realizando o qRT-PCR do gene Rv1258c. RT-PCR foi realizada com RNA previamente isolado de M. tuberculosis CDC1551, a fim de verificar se o Rv1255c, Rv1256c, Rv1257c e Rv1258c genes estão todos ligados em um transcrito. A cepa M. bovis BCG, expressando uma proteína fluorescente vermelha sob o controle do promotor do Rv1258c (pVVTapPro) foi cultivado na presença de estreptomicina e glicina para verificar a indução deste promotor. O qRT-PCR demonstrou uma regulação negativa do gene Rv1258c na cepa sobre expressando Rv1257c, mas não foi observada diferença na cepa sobre expressando Rv1255c em comparação com o controle. Um aumento de fluorescência foi observada na cepa contendo o promotor da Tap na presença de 1 mg/L de estreptomicina, em comparação com o controle. Uma indução de duas vezes do gene Tap foi também observada na presença de glicina 1,6 mM. Os resultados sugerem um papel para o gene Rv1257c na regulação da expressão Tap. Além disso, a indução da Tap por estreptomicina apoia a importância desta proteína na multidroga-resistência de M. tuberculosis. O efeito da glicina no promotor da Tap sugere um papel fisiológico de detoxificação celular para essa bomba de efluxo. Em conclusão este estudo reforça a necessidade de se obter um conhecimento mais aprofundado a respeito do operon da proteína Tap e sua regulação, de maneira a auxiliar no desenvolvimento de inibidores dessa bomba de efluxo e possivelmente de outros mecanismos de resistência induzida.application/pdfporUniversidade Federal de PelotasPrograma de Pós-Graduação em BiotecnologiaUFPelBRBiotecnologiaTuberculosisDrug resistanceEffluxTuberculosisResistênciaEfluxoCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMALCompostos indutores e genes regulando a expressão da bomba de efluxo Tap em Mycobacterium bovis BCGInductors and genes that regulate expression of the Tap efflux pump in Mycobacterium bovis BCGinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttp://lattes.cnpq.br/6277555029608826http://lattes.cnpq.br/4577337828511155Dellagostin, Odir Antôniohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723107D9Silva, Pedro Eduardo Almeida daFelix, Carolina Rodriguesinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFPel - Guaiacainstname:Universidade Federal de Pelotas (UFPEL)instacron:UFPELORIGINALdissertacao_carolina_felix.pdfapplication/pdf7826200http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1222/1/dissertacao_carolina_felix.pdf4432d1e031f9b3b4bdbc3946dbb901e0MD51open accessTEXTdissertacao_carolina_felix.pdf.txtdissertacao_carolina_felix.pdf.txtExtracted Texttext/plain74084http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1222/2/dissertacao_carolina_felix.pdf.txt4f7233e1ef9b767c6e306b742b2a1068MD52open accessTHUMBNAILdissertacao_carolina_felix.pdf.jpgdissertacao_carolina_felix.pdf.jpgGenerated Thumbnailimage/jpeg1748http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1222/3/dissertacao_carolina_felix.pdf.jpgd04b23d4a2439dfac0f9fd956254ddf0MD53open access123456789/12222019-08-23 10:06:05.99open accessoai:guaiaca.ufpel.edu.br:123456789/1222Repositório InstitucionalPUBhttp://repositorio.ufpel.edu.br/oai/requestrippel@ufpel.edu.br || repositorio@ufpel.edu.br || aline.batista@ufpel.edu.bropendoar:2019-08-23T13:06:05Repositório Institucional da UFPel - Guaiaca - Universidade Federal de Pelotas (UFPEL)false |
dc.title.por.fl_str_mv |
Compostos indutores e genes regulando a expressão da bomba de efluxo Tap em Mycobacterium bovis BCG |
dc.title.alternative.eng.fl_str_mv |
Inductors and genes that regulate expression of the Tap efflux pump in Mycobacterium bovis BCG |
title |
Compostos indutores e genes regulando a expressão da bomba de efluxo Tap em Mycobacterium bovis BCG |
spellingShingle |
Compostos indutores e genes regulando a expressão da bomba de efluxo Tap em Mycobacterium bovis BCG Felix, Carolina Rodrigues Tuberculosis Drug resistance Efflux Tuberculosis Resistência Efluxo CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMAL |
title_short |
Compostos indutores e genes regulando a expressão da bomba de efluxo Tap em Mycobacterium bovis BCG |
title_full |
Compostos indutores e genes regulando a expressão da bomba de efluxo Tap em Mycobacterium bovis BCG |
title_fullStr |
Compostos indutores e genes regulando a expressão da bomba de efluxo Tap em Mycobacterium bovis BCG |
title_full_unstemmed |
Compostos indutores e genes regulando a expressão da bomba de efluxo Tap em Mycobacterium bovis BCG |
title_sort |
Compostos indutores e genes regulando a expressão da bomba de efluxo Tap em Mycobacterium bovis BCG |
author |
Felix, Carolina Rodrigues |
author_facet |
Felix, Carolina Rodrigues |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/6277555029608826 |
dc.contributor.advisorLattes.por.fl_str_mv |
http://lattes.cnpq.br/4577337828511155 |
dc.contributor.advisor-co1.fl_str_mv |
Dellagostin, Odir Antônio |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723107D9 |
dc.contributor.advisor1.fl_str_mv |
Silva, Pedro Eduardo Almeida da |
dc.contributor.author.fl_str_mv |
Felix, Carolina Rodrigues |
contributor_str_mv |
Dellagostin, Odir Antônio Silva, Pedro Eduardo Almeida da |
dc.subject.eng.fl_str_mv |
Tuberculosis Drug resistance Efflux |
topic |
Tuberculosis Drug resistance Efflux Tuberculosis Resistência Efluxo CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMAL |
dc.subject.por.fl_str_mv |
Tuberculosis Resistência Efluxo |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMAL |
description |
Transport proteins related to drug efflux play an important role not only in the acquisition of drug-resistant phenotypes, but also in virulence of M. tuberculosis. The main objective of this study was to analyze the regulation of genes encoding the protein Rv1258c Tap considering the expression of genes Rv1255c and Rv1257c. RNA was extracted from cultures of M. bovis BCG overexpressing Rv1255c and Rv1257c, as well as a control strain containing the vector pVV16, and qRT-PCR of the Rv1258c gene was performed. RT-PCR was performed using RNA isolated previously from M. tuberculosis CDC1551, in order to verify that Rv1255c, Rv1256c, Rv1257c and Rv1258c genes are all connected to a transcript. The strain M. bovis BCG, expressing a red fluorescent protein under control of the promoter Rv1258c (pVVTapPro) was grown in the presence of streptomycin and glycine to verify the induction of the promoter. The qRT-PCR showed a downregulation of the gene in the strain overexpressing Rv1258c and Rv1257c, but no difference was observed in the strain overexpressing Rv1255c compared with the control. An increase in fluorescence was observed in the strain containing the promoter of the tap in the presence of 1 mg/L of streptomycin in comparison with the control. A two-fold induction of gene Tap was also observed in the presence of 1.6 mM glycine. The results suggest a role for Rv1257c in the regulation Tap expression. Moreover, the induction of Tap by streptomycin supports the importance of this protein in multidrugresistant M. tuberculosis. The effect of glycine on Tap promoter suggest a physiological role in cellular detoxification for this efflux pump. In conclusion, this study reinforces the need to obtain a deeper knowledge about the Tap protein operon and its regulation, in order to assist in the development of inhibitors of this efflux pump and possibly other mechanisms of induced resistance. |
publishDate |
2013 |
dc.date.issued.fl_str_mv |
2013-08-28 |
dc.date.accessioned.fl_str_mv |
2014-08-20T13:32:48Z |
dc.date.available.fl_str_mv |
2014-01-24 2014-08-20T13:32:48Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
FELIX, Carolina Rodrigues. Inductors and genes that regulate expression of the Tap efflux pump in Mycobacterium bovis BCG. 2013. 52 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2013. |
dc.identifier.uri.fl_str_mv |
http://guaiaca.ufpel.edu.br/handle/123456789/1222 |
identifier_str_mv |
FELIX, Carolina Rodrigues. Inductors and genes that regulate expression of the Tap efflux pump in Mycobacterium bovis BCG. 2013. 52 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2013. |
url |
http://guaiaca.ufpel.edu.br/handle/123456789/1222 |
dc.language.iso.fl_str_mv |
por |
language |
por |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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Universidade Federal de Pelotas |
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Programa de Pós-Graduação em Biotecnologia |
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UFPel |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Biotecnologia |
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Universidade Federal de Pelotas |
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