EFFICIENT INDUCTION OF POINT MUTATIONS ALLOWING RECOVERY OF SPECIFIC-LOCUS MUTATIONS IN ZEBRAFISH
EFFICIENT INDUCTION OF POINT MUTATIONS ALLOWING RECOVERY OF SPECIFIC-LOCUS MUTATIONS IN ZEBRAFISH
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DOI：
10.1073/pnas.92.13.5997
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发表时间：
1995-06-20
影响因子：
11.1
通讯作者：
GRUNWALD, DJ
中科院分区：
文献类型：
--
作者：
RILEY, BB;GRUNWALD, DJ
描述了一种极大地提高恢复斑马鱼(Danio Rerio)特定点点突变的效率的技术。用烷基化试剂N-乙基-N-亚硝脲诱变减数分裂后配子，产生嵌合点突变的创始个体。在最佳条件下，每个创始人平均携带10个突变，影响胚胎发育所需的基因。此外，这些创始人中约有2%的人在任何预先指定的色素沉着基因座上传递了新的突变。对新的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-亚硝脲对减数分裂后配子进行诱变后，在特定位点产生的点突变频率比以前在斑马鱼中获得的点突变频率高10-15倍。因此，我们的程序应该极大地促进在任何感兴趣的座位上恢复多个突变等位基因。
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, approximate to 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.