Targeted screening for induced mutations
Targeted screening for induced mutations
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DOI：
10.1038/74542
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发表时间：
2000-04-01
影响因子：
46.9
通讯作者：
Henikoff, S
中科院分区：
文献类型：
--
作者：
McCallum, CM;Comai, L;Henikoff, S
随着大规模序列数据的积累，基因组学研究的重点已经从确定基因结构转向检测基因功能，而这依赖于反向遗传方法论。在这里，我们探索了在拟南芥中筛选化学诱导的靶序列突变的可行性。我们的TILLING(靶向诱导基因组局部损伤)方法结合了甲烷磺酸乙酯(EMS)诱导突变的效率1和变性高效液相色谱(DHPLC)通过异源双链分析检测碱基对变化的能力2。重要的是，这种方法产生了广泛的突变等位基因，快速和自动化，适用于任何可以化学诱变的有机体。
With the accumulation of large-scale sequence data, emphasis in genomics has shifted from determining gene structure to testing gene function, and this relies on reverse genetic methodology. Here we explore the feasibility of screening for chemically induced mutations in target sequences in Arabidopsis thaliana. Our TILLING (Targeting Induced Local Lesions IN Genomes) method combines the efficiency of ethyl methanesulfonate (EMS)-induced mutagenesis 1 with the ability of denaturing high-performance liquid chromatography (DHPLC) to detect base pair changes by heteroduplex analysis 2. Importantly, this method generates a wide range of mutant alleles, is fast and automatable, and is applicable to any organism that can be chemically mutagenized.