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

• 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
• topic: Alkaline gel electrophoresis; Escherichia coli; base excision repair; stannous chloride; DNA strand breaks; reactive oxygen species

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.