Department of Biology, University of Winnipeg, 515 Portage Avenue, Winnipeg, MB R3B 2E9, Canada.
Genome. 2011 Apr;54(4):341-7. doi: 10.1139/g10-120.
The recent completion of genome sequencing of 12 species of Drosophila has provided a powerful resource for hypothesis testing, as well as the development of technical tools. Here we take advantage of genome sequence data from two closely related species of Drosophila, Drosophila simulans and Drosophila sechellia, to quickly identify candidate molecular markers for genotyping based on expected insertion or deletion (indel) differences between species. Out of 64 candidate molecular markers selected along the second and third chromosome of Drosophila, 51 molecular markers were validated using PCR and gel electrophoresis. We found that the 20% error rate was due to sequencing errors in the genome data, although we cannot rule out possible indel polymorphisms. The approach has the advantage of being affordable and quick, as it only requires the use of bioinformatics tools for predictions and a PCR and agarose gel based assay for validation. Moreover, the approach could be easily extended to a wide variety of taxa with the only limitation being the availability of complete or partial genome sequence data.
最近完成的 12 种果蝇的基因组测序为假设检验以及技术工具的开发提供了强大的资源。在这里,我们利用来自两个密切相关的果蝇物种——黑腹果蝇和塞舌尔果蝇的基因组序列数据,根据物种之间预期的插入或缺失(indel)差异,快速识别候选的分子标记用于基因分型。在沿着黑腹果蝇的第二和第三染色体选择的 64 个候选分子标记中,有 51 个分子标记通过 PCR 和凝胶电泳进行了验证。我们发现,20%的错误率归因于基因组数据中的测序错误,尽管我们不能排除可能的 indel 多态性。该方法具有经济实惠和快速的优点,因为它仅需要使用生物信息学工具进行预测,以及 PCR 和琼脂糖凝胶检测进行验证。此外,该方法可以很容易地扩展到各种具有完整或部分基因组序列数据的分类单元,唯一的限制是这些数据的可用性。