Large-scale mapping of mutations affecting zebrafish development.
Large-scale mapping of mutations affecting zebrafish development.
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DOI：
10.1186/1471-2164-8-11
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发表时间：
2007-01-09
期刊：
影响因子：
4.4
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中科院分区：
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在使用诱变剂ENU的斑马鱼中进行的大规模突变筛选已经分离出数百个代表可能的发育控制基因的突变位点。为了实现这种筛查的潜力，有必要对突变进行系统的基因图谱绘制。在这里，我们报告了一项大规模的努力，通过使用微卫星标记的基因组扫描来绘制马克斯·普朗克发育生物学研究所在突变筛选中产生的突变。我们已经选择了一套微卫星标记，并开发了适合高效、高通量基因组扫描的方法和评分标准。我们已经使用这些方法从Tübingen I突变筛选和随后的突变集合筛选中成功地获得了319个突变座位的粗略图谱位置。对于其中的277个，相应的基因还没有确定。80%的被测基因座作图成功。通过比较21个突变和克隆突变的基因位置，我们验证了我们的连锁群分配的正确性，并估计了我们的图谱位置的标准误差约为6 cM。通过获得300多个具有发育表型的斑马鱼基因座的粗略图谱位置，我们已经生成了一个数据集，该数据集不仅对克隆受影响的基因有用，而且还将在未来的筛查中发现具有类似表型的突变的等位基因。此外，这项工作验证了我们的方法对于快速、系统和廉价的斑马鱼突变的微卫星图谱的有效性。
Large-scale mutagenesis screens in the zebrafish employing the mutagen ENU have isolated several hundred mutant loci that represent putative developmental control genes. In order to realize the potential of such screens, systematic genetic mapping of the mutations is necessary. Here we report on a large-scale effort to map the mutations generated in mutagenesis screening at the Max Planck Institute for Developmental Biology by genome scanning with microsatellite markers. We have selected a set of microsatellite markers and developed methods and scoring criteria suitable for efficient, high-throughput genome scanning. We have used these methods to successfully obtain a rough map position for 319 mutant loci from the Tübingen I mutagenesis screen and subsequent screening of the mutant collection. For 277 of these the corresponding gene is not yet identified. Mapping was successful for 80 % of the tested loci. By comparing 21 mutation and gene positions of cloned mutations we have validated the correctness of our linkage group assignments and estimated the standard error of our map positions to be approximately 6 cM. By obtaining rough map positions for over 300 zebrafish loci with developmental phenotypes, we have generated a dataset that will be useful not only for cloning of the affected genes, but also to suggest allelism of mutations with similar phenotypes that will be identified in future screens. Furthermore this work validates the usefulness of our methodology for rapid, systematic and inexpensive microsatellite mapping of zebrafish mutations.