Riley B B, Grunwald D J
Department of Human Genetics, University of Utah, Salt Lake City 84112, USA.
Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):5997-6001. doi: 10.1073/pnas.92.13.5997.
A technique is described that greatly increases the efficiency of recovering specific locus point mutations in zebrafish (Danio rerio). Founder individuals that were mosaic for point mutations were produced by mutagenizing postmeiotic gametes with the alkylating agent N-ethyl-N-nitrosourea. Under optimal conditions, each founder carried an average of 10 mutations affecting genes required for embryogenesis. Moreover, approximately 2% of these founders transmitted new mutations at any prespecified pigmentation locus. Analyses of new pigmentation mutations confirmed that most were likely to be point mutations. Thus, mutagenesis of postmeiotic gametes with N-ethyl-N-nitrosourea yielded frequencies of point mutations at specific loci that were 10- to 15-fold higher than previously achieved in zebrafish. Our procedure should, therefore, greatly facilitate recovery of multiple mutant alleles at any locus of interest.
本文描述了一种可大幅提高在斑马鱼（Danio rerio）中恢复特定基因座点突变效率的技术。通过用烷基化剂N-乙基-N-亚硝基脲诱变减数分裂后的配子，产生了携带点突变嵌合体的奠基个体。在最佳条件下，每个奠基个体平均携带10个影响胚胎发育所需基因的突变。此外，这些奠基个体中约2%在任何预先指定的色素沉着基因座传递了新的突变。对新色素沉着突变的分析证实，大多数可能是点突变。因此，用N-乙基-N-亚硝基脲诱变减数分裂后的配子，在特定基因座产生的点突变频率比斑马鱼此前所达到的高出10至15倍。因此，我们的方法应能极大地促进在任何感兴趣的基因座上恢复多个突变等位基因。
