Kikuno T, Honma M, Ogura S, Mizusawa H, Hayashi M, Sofuni T
Department of Toxicology, Hita Research Laboratories, Chemicals Inspection and Testing Institute, Ohita, Japan.
Mutat Res. 1995 Oct;338(1-6):87-93. doi: 10.1016/0921-8734(95)00014-w.
Using a multi-locus minisatellite Per-6 DNA probe, we performed DNA fingerprint analysis. Chinese hamster lung (CHL) cells were treated with six model chemicals: N-methyl-N'-nitro-N-nitrosoguanidine, mitomycin C, methyl methanesulfonate, furylfuramide, 2-acetylamino-fluorene, and cyclophosphamide, with or without S9 mix. 771 hypoxanthine phosphoribosyltransferase deficient clones (749 from mutagen-treated cells and 22 from untreated cells) and 90 unselected clones from untreated cells were isolated and analyzed. The spontaneous mutation frequency at CHL cell minisatellite loci was 0.31-0.63%. All the chemicals increased mutation frequencies. Almost all mutations localized to the three specific minisatellite loci corresponding to 4.2, 3.8, and 2.4 kb bands, suggesting that these regions are more unstable and susceptible to mutation. DNA fingerprint analysis is a promising technique for detecting mutations at neutral DNA regions, especially recombinational mutations, and may be useful for surveying genetic instability related to heritable defects or aging.
使用多位点小卫星Per-6 DNA探针，我们进行了DNA指纹分析。用六种模型化学品处理中国仓鼠肺（CHL）细胞：N-甲基-N'-硝基-N-亚硝基胍、丝裂霉素C、甲磺酸甲酯、呋喃糠酰胺、2-乙酰氨基芴和环磷酰胺，有或没有S9混合液。分离并分析了771个次黄嘌呤磷酸核糖基转移酶缺陷克隆（749个来自诱变处理的细胞，22个来自未处理的细胞）和90个来自未处理细胞的未选择克隆。CHL细胞小卫星位点的自发突变频率为0.31-0.63%。所有化学品均增加了突变频率。几乎所有突变都定位在对应于4.2、3.8和2.4 kb条带的三个特定小卫星位点，表明这些区域更不稳定且易发生突变。DNA指纹分析是检测中性DNA区域突变，尤其是重组突变的一种有前景的技术，可能有助于调查与遗传性缺陷或衰老相关的遗传不稳定性。
