Mutagenic effects at hprt locus and in minisatellite sequences induced in V79 cells by treatments with UV and methyl-nitro-nitroso guanidine

Mutation Research Letters Pub Date : 1995-12-01 DOI:10.1016/0165-7992(95)90009-8

S. Ogheri, C. Rampazzo, L. Celotti

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引用次数: 11

Abstract

DNA alterations induced in V79 cells treated with UV light or methyl-nitro-nitrosoguanidine were analyzed by the mutagenicity test at the hprt locus and by DNA fingerprint analysis. Treated and control cells were seeded in the presence or absence of 6-thioguanine to determine mutant frequency and cell survival. From clonal cultures of the same cell populations we isolated a number of clones and grew them up individually to obtain appropriate amounts of DNA. High molecular weight DNA was digested with HinfI or HaeIII and hybridized with ³²P-labelled 33.15 multilocus probe. The induction of 6-thioguanine resistant cells depended on the mutagen does. The highest value of mutant frequency obtained was 7475 × 10⁻⁶ (MNNG, 27μM), corresponding to 0.7 percent of clonable cells. DNA fingerprint analysis carried out on the same treated cells showed that DNA rearrangements occurred at minisatellites much more frequently than in transcribed sequences. UV irradiation produced the highest frequency of variation, modifying minisatellite patterns in about 50 percent of the analyzed clones.

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紫外线和甲基-硝基-亚硝基胍对V79细胞hprt位点和小卫星序列的诱变效应

通过hprt位点诱变试验和DNA指纹分析，分析了紫外光和甲基硝基-亚硝基胍对V79细胞的诱导作用。在存在或不存在6-硫鸟嘌呤的情况下播种处理和对照细胞，以确定突变频率和细胞存活率。从相同细胞群的克隆培养中，我们分离出许多克隆，并单独培养，以获得适当数量的DNA。用HinfI或HaeIII酶切高分子量DNA，并用32p标记的33.15多位点探针进行杂交。6-硫鸟嘌呤抗性细胞的诱导取决于诱变原。突变频率的最大值为7475 × 10−6 (MNNG, 27μM)，占克隆细胞的0.7%。对同样处理过的细胞进行的DNA指纹分析表明，DNA重排发生在小卫星上的频率远高于转录序列。紫外线照射产生的变异频率最高，大约50%的被分析克隆的小卫星模式发生了改变。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Mutation Research Letters

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