Characterization of mutations induced by ethyl methanesulfonate, UV, and trimethylpsoralen in the nematode Caenorhabditis elegans
Characterization of mutations induced by ethyl methanesulfonate, UV, and trimethylpsoralen in the nematode Caenorhabditis elegans
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DOI：
10.1006/bbrc.2000.2260
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发表时间：
2000-03-05
影响因子：
3.1
通讯作者：
Mitani, S
中科院分区：
文献类型：
--
作者：
Gengyo-Ando, K;Mitani, S
秀丽隐杆线虫的基因组计划完成。对线虫基因进行破坏，以了解其功能是非常重要和有用的。我们在存在TMP（三甲基补骨脂素）的情况下，用反向遗传学诱变剂EMS（甲磺酸乙酯）、短波长紫外线和长波长紫外线的潜在候选物处理野生型动物。我们通过计数标记unc-22突变的发生来估计正向突变率。我发现TMP/UV的正向突变率可以与EMS相当，但频率比以前提高了一个数量级。接下来，我们分离另一个标记基因ben-1的突变体，并通过PCR和测序检查缺失突变的概率。TMP/UV法仅检出缺失突变，提示TMP/UV法是缺失突变的首选方法。作为一个试点实验，我们可以以比以前高得多的频率分离出实际的缺失突变。(C)北京大学出版社.
The genome project of the nematode Caenorhabditis elegans is completed. It is important and useful to disrupt nematode genes to know their function. We treated wild-type animals with potential candidates for mutagens for reverse genetics, EMS (ethyl methanesulfonate), short-wavelength UV, and long-wavelength UV in the presence of TMP (trimethylpsoralen). We estimated forward mutation rates by counting the occurrence of a marker unc-22 mutation. me found that the forward mutation rate by TMP/UV could be comparable with EMS by improving the frequency one order higher than before. We next isolated mutants of another marker gene ben-1 and examined the probability for the deletion mutations by PCR and sequencing. Deletion mutations were found only by TMP/UV method, which suggested TMP/UV is the choice for deletion mutagenesis among these methods. As a pilot experiment, we could isolate actual deletion mutations at a much higher frequency than previously. (C) 2000 Academic Press.