Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar
Autor(a) principal: | |
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Data de Publicação: | 2007 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFRN |
Texto Completo: | https://repositorio.ufrn.br/jspui/handle/123456789/16780 |
Resumo: | Plants are organisms sessile and because of this they are susceptible to genotoxic effects due to environmental exposure such as light [including ultraviolet (UV)], heat, drought and chemicals agents. Therefore, there are differents pathways in order to detect a lesion and correct. These pathways are not well known in plants. The MutM/Fpg protein is a DNA glycosylase that is responsible for detect and correct oxidative lesions. In the sugarcane genome, it was found two possible cDNAs that had homology to this protein: scMUTM1 and scMUTM2. The aim of this work was to characterize the role of these cDNAs in plants. In order to do this, the expression level after oxidative stress was evaluated by semiquantitative RT-PCR. Another point analyzed in order to obtain the full-length gene, it was to use a sugarcane genomic library that was hybridized with both cDNAs as a probe. It was found two clones that will bought and sequenced. The promoter region was also cloned. It was obtained sequences only for scMUTM2 promoter region. The sequences obtained were divided into six groups. It was found regulatory motifs such as TATA-box, CAAT-box, oxidative stress element response and regulatory regions that response to light. The other point analyzed was to characterize the N-terminal region by PCR constructs. These constructs have deletions at 5 region. These sequences were introduce into Escherichia coli wild type strain (CC104) and double mutant (CC104mutMmutY). The results showed that proteins with deletions of scMUTM1 N-terminal region were able to complement the Fpg and MutY-glycosylase deficiency in CC104 mutMmutY reducing the spontaneous mutation frequency |
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Medeiros, Viviane Katielly Silvahttp://lattes.cnpq.br/0635169659175207http://lattes.cnpq.br/4808910380593455Lima, Lucymara Fassarela Agnezhttp://lattes.cnpq.br/1083882171718362Machado, Carlos Renatohttp://lattes.cnpq.br/6306925202374274Scortecci, Kátia Castanho2014-12-17T15:18:13Z2007-07-182014-12-17T15:18:13Z2007-02-23MEDEIROS, Viviane Katielly Silva. Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar. 2007. 4 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2007.https://repositorio.ufrn.br/jspui/handle/123456789/16780Plants are organisms sessile and because of this they are susceptible to genotoxic effects due to environmental exposure such as light [including ultraviolet (UV)], heat, drought and chemicals agents. Therefore, there are differents pathways in order to detect a lesion and correct. These pathways are not well known in plants. The MutM/Fpg protein is a DNA glycosylase that is responsible for detect and correct oxidative lesions. In the sugarcane genome, it was found two possible cDNAs that had homology to this protein: scMUTM1 and scMUTM2. The aim of this work was to characterize the role of these cDNAs in plants. In order to do this, the expression level after oxidative stress was evaluated by semiquantitative RT-PCR. Another point analyzed in order to obtain the full-length gene, it was to use a sugarcane genomic library that was hybridized with both cDNAs as a probe. It was found two clones that will bought and sequenced. The promoter region was also cloned. It was obtained sequences only for scMUTM2 promoter region. The sequences obtained were divided into six groups. It was found regulatory motifs such as TATA-box, CAAT-box, oxidative stress element response and regulatory regions that response to light. The other point analyzed was to characterize the N-terminal region by PCR constructs. These constructs have deletions at 5 region. These sequences were introduce into Escherichia coli wild type strain (CC104) and double mutant (CC104mutMmutY). The results showed that proteins with deletions of scMUTM1 N-terminal region were able to complement the Fpg and MutY-glycosylase deficiency in CC104 mutMmutY reducing the spontaneous mutation frequencyAs plantas são organismos particularmente susceptíveis aos efeitos genotóxicos devido a sua constante exposição a agentes do meio ambiente tais como a luz [incluindo a ultravioleta (UV)], o calor, a seca e diferentes substâncias químicas. Entretanto, existem diferentes mecanismos de reparo para detectar estas lesões e corrigi-las. Entretanto, em plantas estas vias são pouco compreendidas. Uma das vias é a por excisão de bases (BER). A proteína MUTM/FPG é uma DNA glicosilase da via BER que corrige danos oxidativos. No genoma da cana-de-açúcar foram encontrados dois possíveis cDNAs que possuem homologia de seqüência a esta proteína: scMUTM1 e scMUTM2. O objetivo deste trabalho foi caracterizar o papel destes genes em plantas. Para isto, o nível de expressão dos genes após estresse oxidativo foi avaliado através de RT-PCR semiquantitativo. Para obter a sequência completa dos gene, uma biblioteca de DNA genômico de cana-de-açúcar foi hibridizada com sondas radioativas para ambos cDNAs, encontrando-se dois clones em potencial. A região promotora foi clonada, seqüenciada e analisada. Dentro das seqüências obtidas para a região promotora de scMUTM2, estas podem ser divididas em seis grupos. Os motivos regulatórios foram identificados, tais como o TATA-box, CAAT-box, elementos de resposta ao estresse oxidativo e elementos de resposta à luz. Um outro ponto de interesse neste trabalho foi a caracterização da região N-terminal da proteína scMUTM1. Para isto, foram realizadas algumas construções via PCR. Estas construções foram posteriormente introduzidas em cepas selvagem (CC104) e duplo mutante (CC104mutMmutY) de Escherichia coli e foram realizados ensaios de complementação de modo a avaliar a importância de cada região para o reconhecimento e correção das lesões. Os resultados mostraram que as proteínas com deleções na porção N-terminal de scMUTM1 são capazes de complementar a deficiência de Fpg and MutY-glicosilase na bactéria CC104 mutMmutY reduzindo assim a freqüência de mutação espontâneaConselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal do Rio Grande do NortePrograma de Pós-Graduação em Genética e Biologia MolecularUFRNBRGenética e Biologia MolecularCana-de-açúcarDNA glicosilaseReparo de DNASugarcaneDNA glycosylaseDNA repairCNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA MOLECULAR E DE MICROORGANISMOSCaracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcarinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRNinstname:Universidade Federal do Rio Grande do Norte (UFRN)instacron:UFRNORIGINALVivianeKM.pdfapplication/pdf34009https://repositorio.ufrn.br/bitstream/123456789/16780/1/VivianeKM.pdf80f5f4645fd6e0b6573613b53e2fcf67MD51TEXTVivianeKM.pdf.txtVivianeKM.pdf.txtExtracted texttext/plain4880https://repositorio.ufrn.br/bitstream/123456789/16780/6/VivianeKM.pdf.txtb0e778e377514dacd633f12fefbfc572MD56THUMBNAILVivianeKM.pdf.jpgVivianeKM.pdf.jpgIM Thumbnailimage/jpeg2840https://repositorio.ufrn.br/bitstream/123456789/16780/7/VivianeKM.pdf.jpg653278b39851a0c091d1ee626984cd2eMD57123456789/167802017-11-04 06:01:09.588oai:https://repositorio.ufrn.br:123456789/16780Repositório de PublicaçõesPUBhttp://repositorio.ufrn.br/oai/opendoar:2017-11-04T09:01:09Repositório Institucional da UFRN - Universidade Federal do Rio Grande do Norte (UFRN)false |
dc.title.por.fl_str_mv |
Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar |
title |
Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar |
spellingShingle |
Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar Medeiros, Viviane Katielly Silva Cana-de-açúcar DNA glicosilase Reparo de DNA Sugarcane DNA glycosylase DNA repair CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA MOLECULAR E DE MICROORGANISMOS |
title_short |
Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar |
title_full |
Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar |
title_fullStr |
Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar |
title_full_unstemmed |
Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar |
title_sort |
Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar |
author |
Medeiros, Viviane Katielly Silva |
author_facet |
Medeiros, Viviane Katielly Silva |
author_role |
author |
dc.contributor.authorID.por.fl_str_mv |
|
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/0635169659175207 |
dc.contributor.advisorID.por.fl_str_mv |
|
dc.contributor.advisorLattes.por.fl_str_mv |
http://lattes.cnpq.br/4808910380593455 |
dc.contributor.referees1.pt_BR.fl_str_mv |
Lima, Lucymara Fassarela Agnez |
dc.contributor.referees1ID.por.fl_str_mv |
|
dc.contributor.referees1Lattes.por.fl_str_mv |
http://lattes.cnpq.br/1083882171718362 |
dc.contributor.referees2.pt_BR.fl_str_mv |
Machado, Carlos Renato |
dc.contributor.referees2ID.por.fl_str_mv |
|
dc.contributor.referees2Lattes.por.fl_str_mv |
http://lattes.cnpq.br/6306925202374274 |
dc.contributor.author.fl_str_mv |
Medeiros, Viviane Katielly Silva |
dc.contributor.advisor1.fl_str_mv |
Scortecci, Kátia Castanho |
contributor_str_mv |
Scortecci, Kátia Castanho |
dc.subject.por.fl_str_mv |
Cana-de-açúcar DNA glicosilase Reparo de DNA |
topic |
Cana-de-açúcar DNA glicosilase Reparo de DNA Sugarcane DNA glycosylase DNA repair CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA MOLECULAR E DE MICROORGANISMOS |
dc.subject.eng.fl_str_mv |
Sugarcane DNA glycosylase DNA repair |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA MOLECULAR E DE MICROORGANISMOS |
description |
Plants are organisms sessile and because of this they are susceptible to genotoxic effects due to environmental exposure such as light [including ultraviolet (UV)], heat, drought and chemicals agents. Therefore, there are differents pathways in order to detect a lesion and correct. These pathways are not well known in plants. The MutM/Fpg protein is a DNA glycosylase that is responsible for detect and correct oxidative lesions. In the sugarcane genome, it was found two possible cDNAs that had homology to this protein: scMUTM1 and scMUTM2. The aim of this work was to characterize the role of these cDNAs in plants. In order to do this, the expression level after oxidative stress was evaluated by semiquantitative RT-PCR. Another point analyzed in order to obtain the full-length gene, it was to use a sugarcane genomic library that was hybridized with both cDNAs as a probe. It was found two clones that will bought and sequenced. The promoter region was also cloned. It was obtained sequences only for scMUTM2 promoter region. The sequences obtained were divided into six groups. It was found regulatory motifs such as TATA-box, CAAT-box, oxidative stress element response and regulatory regions that response to light. The other point analyzed was to characterize the N-terminal region by PCR constructs. These constructs have deletions at 5 region. These sequences were introduce into Escherichia coli wild type strain (CC104) and double mutant (CC104mutMmutY). The results showed that proteins with deletions of scMUTM1 N-terminal region were able to complement the Fpg and MutY-glycosylase deficiency in CC104 mutMmutY reducing the spontaneous mutation frequency |
publishDate |
2007 |
dc.date.available.fl_str_mv |
2007-07-18 2014-12-17T15:18:13Z |
dc.date.issued.fl_str_mv |
2007-02-23 |
dc.date.accessioned.fl_str_mv |
2014-12-17T15:18:13Z |
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 |
MEDEIROS, Viviane Katielly Silva. Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar. 2007. 4 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2007. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufrn.br/jspui/handle/123456789/16780 |
identifier_str_mv |
MEDEIROS, Viviane Katielly Silva. Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar. 2007. 4 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2007. |
url |
https://repositorio.ufrn.br/jspui/handle/123456789/16780 |
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por |
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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 do Rio Grande do Norte |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Genética e Biologia Molecular |
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UFRN |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Genética e Biologia Molecular |
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Universidade Federal do Rio Grande do Norte |
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reponame:Repositório Institucional da UFRN instname:Universidade Federal do Rio Grande do Norte (UFRN) instacron:UFRN |
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