Application of the single cell gel electrophoresis on yeast cells.
In the present paper, we have applied the single cell gel electrophoresis (SCGE) assay on yeast cells treating Saccharomyces cerevisiae cells with hydrogen peroxide and methyl methanesulfonate (MMS), two DNA damaging agents. In order to overcome the problem with the yeast cell wall that prevented DNA to be extended by the electric field, we disintegrated the cell wall after embedding the cells in agarose. A characteristic picture of comets with residual nuclei and tails was observed and the length of the comet tails was dependent on the concentration of the damaging agents. Yeast cells developed comets at concentrations at least 10 times lower than the concentrations at which comets begin to appear in mammalian cells after treatment with the two genotoxic agents. The higher sensitivity of the yeast comet assay and the fact that S. cerevisiae is one of the most thoroughly studied and easy to work with eukaryotic model system suggest that the proposed method could be an useful tool for investigation of the DNA damaging activity of potential genotoxins.
Miloshev
George
G
Institute of Molecular Biology, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria. gam@obzor.bio21.bas.bg
Mihaylov
Ivailo
I
Anachkova
Boyka
B
D020552:Comet Assay; D004247:DNA / Q000187:drug effects*; D004249:DNA Damage; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D004247:DNA