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

Brazilian Archives of Biology and Technology

José Carlos Pelielo De Mattos¹  Ellen Serri Da Motta¹  Márcia Betania Nunes De Oliveira¹  Flávio José Da Silva Dantas¹  Adriano Caldeira De Araujo¹ 

[1] ,Universidade do Estado do Rio de Janeiro Departamento de Biofísica e Biometria Laboratório de Radio e FotobiologiaRio de Janeiro RJ ,Brasil

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

DOI  :  10.1590/S1516-89132008000700020

来源: SciELO

【 摘 要 】

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.

【 授权许可】

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