Herron Bruce J, Lu Weining, Rao Cherie, Liu Shanming, Peters Heiko, Bronson Roderick T, Justice Monica J, McDonald J David, Beier David R
Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Nat Genet. 2002 Feb;30(2):185-9. doi: 10.1038/ng812. Epub 2002 Jan 2.
Treatment with N-ethyl-N-nitrosourea (ENU) efficiently generates single-nucleotide mutations in mice. Along with the renewed interest in this approach, much attention has been given recently to large screens with broad aims; however, more finely focused studies have proven very productive as well. Here we show how mutagenesis together with genetic mapping can facilitate the rapid characterization of recessive loci required for normal embryonic development. We screened third-generation progeny of mutagenized mice at embryonic day (E) 18.5 for abnormalities of organogenesis. We ascertained 15 monogenic mutations in the 54 families that were comprehensively analyzed. We carried out the experiment as an outcross, which facilitated the genetic mapping of the mutations by haplotype analysis. We mapped seven of the mutations and identified the affected locus in two lines. Using a hierarchical approach, it is possible to maximize the efficiency of this analysis so that it can be carried out easily with modest infrastructure and resources.
用N-乙基-N-亚硝基脲（ENU）处理可在小鼠中高效产生单核苷酸突变。随着对该方法重新产生兴趣，近来人们对具有广泛目标的大规模筛选给予了很多关注；然而，更具针对性的精细研究也已证明非常有成效。在这里，我们展示了诱变与基因定位如何能够促进对正常胚胎发育所需的隐性基因座的快速表征。我们在胚胎第18.5天（E18.5）筛选诱变小鼠的第三代后代，以寻找器官发生异常情况。在全面分析的54个家系中，我们确定了15个单基因突 变。我们以杂交方式进行实验，这通过单倍型分析促进了突变的基因定位。我们对其中7个突变进行了定位，并在两个品系中确定了受影响的基因座。使用分层方法，可以最大限度地提高这种分析的效率，从而能够在适度的基础设施和资源条件下轻松开展分析。
