Ehlers J, Tosch M, AlBaz I, Lochmann E R
Institut für Biochemie und Molekularbiologie, Freie Universität Berlin, Germany.
Ecotoxicol Environ Saf. 1991 Oct;22(2):133-8. doi: 10.1016/0147-6513(91)90053-r.
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.
我们提出了一种快速评估有毒化学物质对酵母染色体损伤的方法。该方法采用以下步骤：将酵母细胞与化学物质一起孵育，在琼脂糖基质中制备DNA，通过脉冲场凝胶电泳将染色体大小的DNA分子分离成可重复的条带模式，以及通过光密度测定法定量染色体条带的强度。酿酒酵母细胞已用N-甲基-N'-硝基-N-亚硝基胍（MNNG）和顺式二氯二氨合铂（II）（顺铂）处理，已知这两种物质都会与DNA相互作用，还使用了三氯乙烯（TCE），尚未发现其在酵母中具有这种作用。用MNNG和顺铂处理细胞会导致条带模式强度受损，受损程度取决于所施加化学物质的浓度。对于TCE，未观察到类似的效果。该方法作为一种筛选测试，将有助于评估有毒化学物质的生物危害。
