Caracterização dos genes scMUTM1 e scMUTM2 de cana-de-açúcar

Detalhes bibliográficos
Ano de defesa: 2007
Autor(a) principal: Medeiros, Viviane Katielly Silva
Orientador(a): Scortecci, Kátia Castanho
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal do Rio Grande do Norte
Programa de Pós-Graduação: Programa de Pós-Graduação em Genética e Biologia Molecular
Departamento: Genética e Biologia Molecular
País: BR
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: 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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spelling 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
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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.
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dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Genética e Biologia Molecular
publisher.none.fl_str_mv Universidade Federal do Rio Grande do Norte
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