Alkaline gel electrophoresis assay to detect DNA strand breaks and repair mechanisms in Escherichia coli
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Outros Autores: | , , , |
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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Brazilian Archives of Biology and Technology |
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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 |
_version_ |
1750318272681607168 |