Lehrbach Nicolas J, Ji Fei, Sadreyev Ruslan
Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts.
Department of Genetics, Harvard Medical School, Boston, Massachusetts.
Curr Protoc Mol Biol. 2017 Jan 5;117:7.29.1-7.29.12. doi: 10.1002/cpmb.27.
Forward genetic analysis using chemical mutagenesis in model organisms is a powerful tool for investigation of molecular mechanisms in biological systems. In the nematode, Caenorhabditis elegans, mutagenesis screens using ethyl methanesulfonate (EMS) have led to important insights into genetic control of animal development and physiology. A major bottleneck to this approach is identification of the causative mutation underlying a phenotype of interest. In the past, this has required time-consuming genetic mapping experiments. More recently, next-generation sequencing technologies have allowed development of new methods for rapid mapping and identification of EMS-induced lesions. In this unit we describe a protocol to map and identify EMS-induced mutations in C. elegans. © 2017 by John Wiley & Sons, Inc.
在模式生物中利用化学诱变进行正向遗传学分析,是研究生物系统分子机制的有力工具。在线虫秀丽隐杆线虫中,使用甲磺酸乙酯(EMS)进行诱变筛选,已使人们对动物发育和生理学的遗传控制有了重要认识。这种方法的一个主要瓶颈是确定感兴趣表型背后的致病突变。过去,这需要耗时的遗传定位实验。最近,新一代测序技术使得开发快速定位和鉴定EMS诱导损伤的新方法成为可能。在本单元中,我们描述了一种在秀丽隐杆线虫中定位和鉴定EMS诱导突变的方案。© 2017约翰威立父子公司。