摘要
Mutagenesis screening, in which heritable traits are isolated following damage to the genome, is a powerful approach for investigating gene function. Among vertebrate model organisms, the zebrafish (Danio rerio) is ideally suited to mutagenesis screens. The success of large-scale screens is dependent on the way in which changes are identified. The type of damage induced is also pivotal. Single base coding region deletions and insertions are suited to abolition of gene function whilst inducing a small physical alteration to the genome. Such mutations are not commonly found following mutagenesis schemes reported to date. Here, we show that an acridine mutagen, ICR191, which in other model organisms frequently induces single base deletions and insertions, is mutagenic in zebrafish. ICR191 induces hallmark phenotypes associated with genetic damage in treated embryos. Alterations are heritable. Offspring of mutagenised fish had mutations in a marker gene and were found to produce offspring with abnormal development. Using an adaptation of a molecular mutation detection method, fluorescent arbitrary primed PCR, we identified an induced alteration directly. The estimated frequency of induced mutations was sufficiently high to make it feasible to employ this approach for mutagenesis screening.
摘要译文
诱变筛选，其遗传性状在损伤基因组后被分离，是调查基因功能的强大方法。在脊椎动物模型生物体中，斑马鱼（Danio rerio）非常适合诱变筛选。大屏幕的成功取决于识别变化的方式。所造成的损害类型也是至关重要的。单碱基编码区缺失和插入适合于消除基因功能，同时诱导对基因组的小的物理改变。迄今报道的诱变方案之后，这种突变通常不被发现。在这里，我们显示吖啶诱变剂，ICR191，在其他模型生物体中经常诱导单碱基缺失和插入，在斑马鱼中是致突变的。 ICR191诱导与处理的胚胎遗传损伤相关的特征表型。改变是可遗传的。诱变鱼的后代在标记基因中具有突变，发现产生异常发育的后代。使用分子突变检测方法，荧光任意引物PCR，我们直接鉴定诱导的改变。诱导突变的估计频率足够高，以便采用这种方法进行诱变筛选。
Richard Hampson; Simon M. Hughes. Acridine mutagenesis of zebrafish (Danio rerio)[J]. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 2003,525(1-2): 1–9