Rapid estimation of chromosomal damage in yeast due to the effects of environmental chemicals using pulsed field gel electrophoresis.
We present a procedure to rapidly estimate the damage to yeast chromosomes by toxic chemicals. This procedure employs the following steps: incubation of yeast cells with the chemicals, DNA preparation in an agarose matrix, separation of chromosome-sized DNA molecules into reproducible band patterns by pulsed field gel electrophoresis, and quantification of the intensity of chromosomal bands by densitometry. Saccharomyces cerevisiae cells have been treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and cis-Platinum(II) diamminedichloride (cisPT), both of which are known to interact with DNA, and trichlorethylen (TCE), for which such an effect has not been shown in yeast. Treatment of cells with MNNG and cisPt led to an impairment of the intensity of the band pattern to an extent dependent on the concentration of the chemicals applied. For TCE a similar effect could not be discerned. This procedure will be useful as a screening test for the estimation of the biological hazards of toxic chemicals.
Ehlers
J
J
Institut für Biochemie und Molekularbiologie, Freie Universität Berlin, Germany.
Tosch
M
M
AlBaz
I
I
Lochmann
E R
ER
D015825:Chromosomes, Fungal / Q000187:drug effects*; D002945:Cisplatin / Q000633:toxicity; D004271:DNA, Fungal / Q000187:drug effects / Q000302:isolation & purification; D016521:Electrophoresis, Gel, Pulsed-Field; D008769:Methylnitronitrosoguanidine / Q000633:toxicity; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000648:ultrastructure*; D014241:Trichloroethylene / Q000633:toxicity
D004271:DNA, Fungal; D008769:Methylnitronitrosoguanidine; D014241:Trichloroethylene; D002945:Cisplatin