Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli

Detalhes bibliográficos
Autor(a) principal: Mattos,José Carlos Pelielo de
Data de Publicação: 2008
Outros Autores: Motta,Ellen Serri da, Oliveira,Márcia Betania Nunes de, Dantas,Flávio José da Silva, Araujo,Adriano Caldeira de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Archives of Biology and Technology
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132008000700020
Resumo: Reactive oxygen species (ROS) can induce lesions in different cellular targets, including DNA. Stannous chloride (SnCl2) is a ROS generator, leading to lethality in Escherichia coli (E. coli), with the base excision repair (BER) mechanism playing a role in this process. Many techniques have been developed to detect genotoxicity, as comet assay, in eukaryotic cells, and plasmid DNA agarose gel electrophoresis. In this study, an adaptation of the alkaline gel electrophoresis method was carried out to ascertain the induction of strand breaks by SnCl2 in bacterial DNA, from E. coli BER mutants, and its repair pathway. Results obtained show that SnCl2 was able to induce DNA strand breaks in all strains tested. Moreover, endonuclease IV and exonuclease III play a role in DNA repair. On the whole, data has shown that the alkaline gel electrophoresis assay could be used both for studying DNA strand breaks induction and for associated repair mechanisms.
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spelling Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coliAlkaline gel electrophoresisEscherichia colibase excision repairstannous chlorideDNA strand breaksreactive oxygen speciesReactive oxygen species (ROS) can induce lesions in different cellular targets, including DNA. Stannous chloride (SnCl2) is a ROS generator, leading to lethality in Escherichia coli (E. coli), with the base excision repair (BER) mechanism playing a role in this process. Many techniques have been developed to detect genotoxicity, as comet assay, in eukaryotic cells, and plasmid DNA agarose gel electrophoresis. In this study, an adaptation of the alkaline gel electrophoresis method was carried out to ascertain the induction of strand breaks by SnCl2 in bacterial DNA, from E. coli BER mutants, and its repair pathway. Results obtained show that SnCl2 was able to induce DNA strand breaks in all strains tested. Moreover, endonuclease IV and exonuclease III play a role in DNA repair. On the whole, data has shown that the alkaline gel electrophoresis assay could be used both for studying DNA strand breaks induction and for associated repair mechanisms.Instituto de Tecnologia do Paraná - Tecpar2008-12-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132008000700020Brazilian Archives of Biology and Technology v.51 n.spe 2008reponame:Brazilian Archives of Biology and Technologyinstname:Instituto de Tecnologia do Paraná (Tecpar)instacron:TECPAR10.1590/S1516-89132008000700020info:eu-repo/semantics/openAccessMattos,José Carlos Pelielo deMotta,Ellen Serri daOliveira,Márcia Betania Nunes deDantas,Flávio José da SilvaAraujo,Adriano Caldeira deeng2009-02-20T00:00:00Zoai:scielo:S1516-89132008000700020Revistahttps://www.scielo.br/j/babt/https://old.scielo.br/oai/scielo-oai.phpbabt@tecpar.br||babt@tecpar.br1678-43241516-8913opendoar:2009-02-20T00:00Brazilian Archives of Biology and Technology - Instituto de Tecnologia do Paraná (Tecpar)false
dc.title.none.fl_str_mv Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli
title Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli
spellingShingle Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli
Mattos,José Carlos Pelielo de
Alkaline gel electrophoresis
Escherichia coli
base excision repair
stannous chloride
DNA strand breaks
reactive oxygen species
title_short Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli
title_full Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli
title_fullStr Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli
title_full_unstemmed Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli
title_sort Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli
author Mattos,José Carlos Pelielo de
author_facet Mattos,José Carlos Pelielo de
Motta,Ellen Serri da
Oliveira,Márcia Betania Nunes de
Dantas,Flávio José da Silva
Araujo,Adriano Caldeira de
author_role author
author2 Motta,Ellen Serri da
Oliveira,Márcia Betania Nunes de
Dantas,Flávio José da Silva
Araujo,Adriano Caldeira de
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Mattos,José Carlos Pelielo de
Motta,Ellen Serri da
Oliveira,Márcia Betania Nunes de
Dantas,Flávio José da Silva
Araujo,Adriano Caldeira de
dc.subject.por.fl_str_mv Alkaline gel electrophoresis
Escherichia coli
base excision repair
stannous chloride
DNA strand breaks
reactive oxygen species
topic Alkaline gel electrophoresis
Escherichia coli
base excision repair
stannous chloride
DNA strand breaks
reactive oxygen species
description Reactive oxygen species (ROS) can induce lesions in different cellular targets, including DNA. Stannous chloride (SnCl2) is a ROS generator, leading to lethality in Escherichia coli (E. coli), with the base excision repair (BER) mechanism playing a role in this process. Many techniques have been developed to detect genotoxicity, as comet assay, in eukaryotic cells, and plasmid DNA agarose gel electrophoresis. In this study, an adaptation of the alkaline gel electrophoresis method was carried out to ascertain the induction of strand breaks by SnCl2 in bacterial DNA, from E. coli BER mutants, and its repair pathway. Results obtained show that SnCl2 was able to induce DNA strand breaks in all strains tested. Moreover, endonuclease IV and exonuclease III play a role in DNA repair. On the whole, data has shown that the alkaline gel electrophoresis assay could be used both for studying DNA strand breaks induction and for associated repair mechanisms.
publishDate 2008
dc.date.none.fl_str_mv 2008-12-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132008000700020
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132008000700020
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S1516-89132008000700020
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Instituto de Tecnologia do Paraná - Tecpar
publisher.none.fl_str_mv Instituto de Tecnologia do Paraná - Tecpar
dc.source.none.fl_str_mv Brazilian Archives of Biology and Technology v.51 n.spe 2008
reponame:Brazilian Archives of Biology and Technology
instname:Instituto de Tecnologia do Paraná (Tecpar)
instacron:TECPAR
instname_str Instituto de Tecnologia do Paraná (Tecpar)
instacron_str TECPAR
institution TECPAR
reponame_str Brazilian Archives of Biology and Technology
collection Brazilian Archives of Biology and Technology
repository.name.fl_str_mv Brazilian Archives of Biology and Technology - Instituto de Tecnologia do Paraná (Tecpar)
repository.mail.fl_str_mv babt@tecpar.br||babt@tecpar.br
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