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紫外线和甲基硝基亚硝基胍处理V79细胞后在次黄嘌呤磷酸核糖转移酶基因座和小卫星序列中诱导的诱变效应。

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

作者信息

Ogheri S, Rampazzo C, Celotti L

出版信息

Mutat Res. 1995 Dec;348(4):193-9. doi: 10.1016/0165-7992(95)90009-8.

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 dose. The highest value of mutant frequency obtained was 7475 x 10(-6) (MNNG, 27 microM), corresponding to 0.7 percent of clonable cells. DNA fingerprint analysis carried 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.

摘要

通过次黄嘌呤磷酸核糖转移酶(hprt)基因座的致突变性试验和DNA指纹分析,对用紫外线或甲基硝基亚硝基胍处理的V79细胞中诱导的DNA改变进行了分析。在有或没有6-硫鸟嘌呤的情况下接种处理过的细胞和对照细胞,以确定突变频率和细胞存活率。从相同细胞群体的克隆培养物中,我们分离出许多克隆并单独培养,以获得适量的DNA。用HinfI或HaeIII消化高分子量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剂剂量。获得的最高突变频率值为7475×10^(-6)(甲基硝基亚硝基胍,27微摩尔),相当于可克隆细胞的0.7%。对相同处理细胞进行的DNA指纹分析表明,微卫星处的DNA重排比转录序列中更频繁地发生。紫外线照射产生的变异频率最高,约50%的分析克隆中的微卫星模式发生了改变。

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