Efficient recovery of ENU-induced mutations from the zebrafish germline.
Efficient recovery of ENU-induced mutations from the zebrafish germline.
复制标题
从斑马鱼种系中有效恢复 ENU 诱导的突变。
DOI：
10.1093/genetics/136.4.1401
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发表时间：
1994
期刊：
影响因子：
3.3
通讯作者：
Driever,W
中科院分区：
文献类型：
--
作者：
Solnica-Krezel,L;Schier,AF;Driever,W
研究了甲基磺酸乙酯（EMS）和n -乙基-n -亚硝基脲（ENU）两种化学诱变剂对斑马鱼（Brachydanio rerio）精子发生不同阶段突变的诱导效率。EMS和ENU在减数分裂后的生殖细胞中诱导突变的几率都很高，正如白化突变F1后代镶嵌的发生率所表明的那样。然而，对于减数分裂前生殖细胞，只有ENU被发现是一种有效的诱变原，这表明了在四个不同的色素沉着位点的非马赛克突变子代的频率。研究了几种不同治疗次数或ENU浓度的致突变性方案，以获得致突变性和毒性之间的最佳比例。对于两种最具诱变性的方案：3mm ENU 4 × 1小时和3mm ENU 6 × 1小时，每个基因座每个配子独立突变的最小估计频率为0.9-1.3 × 10（-3）。我们证明了ENU诱导的胚胎致死突变可以传递给后代，并且可以在F2筛选中恢复。特异位点突变的平均频率为1.1 X 10(-3)，相当于每个突变基因组约1.7个胚胎致死突变。ENU可实现的高突变率允许快速识别涉及斑马鱼发育各个方面的大量基因。
We studied the efficiency with which two chemical mutagens, ethyl methanesulfonate (EMS) and N-ethyl-N-nitrosourea (ENU) can induce mutations at different stages of spermatogenesis in zebrafish (Brachydanio rerio). Both EMS and ENU induced mutations at high rates in post-meiotic germ cells, as indicated by the incidence of F1 progeny mosaic for the albino mutation. For pre-meiotic germ cells, however, only ENU was found to be an effective mutagen, as indicated by the frequencies of non-mosaic mutant progeny at four different pigmentation loci. Several mutagenic regimens that varied in either the number of treatments or the concentration of ENU were studied to achieve an optimal ratio between the mutagenicity and toxicity. For the two most mutagenic regimens: 4 x 1 hr in 3 mM ENU and 6 x 1 hr in 3 mM ENU, the minimum estimate of frequencies of independent mutations per locus per gamete was 0.9-1.3 X 10(-3). We demonstrate that embryonic lethal mutations induced with ENU were transmitted to offspring and that they could be recovered in an F2 screen. An average frequency of specific-locus mutations of 1.1 X 10(-3) corresponded to approximately 1.7 embryonic lethal mutations per single mutagenized genome. The high rates of mutations achievable with ENU allow for rapid identification of large numbers of genes involved in a variety of aspects of zebrafish development.