Large-scale discovery of induced point mutations with high-throughput TILLING
Large-scale discovery of induced point mutations with high-throughput TILLING
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DOI：
10.1101/gr.977903
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发表时间：
2003-03-01
期刊：
影响因子：
7
通讯作者：
Henikoff, S
中科院分区：
文献类型：
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作者：
Till, BJ;Reynolds, SH;Henikoff, S
TILLING ((T) bar targeting下(I) bar induced下(L) bar ocal下（L) bar esions下(G) bar基因组下）是一种通用的反向遗传策略，它在感兴趣的基因中提供一系列诱导点突变的等位基因。高通量TILLING允许快速和低成本地发现化学诱变个体群体中的诱导点突变。由于化学诱变的广泛应用，TILLING的突变检测仅依赖于PCR产物的足够产量，因此TILLING可以应用于大多数生物。我们已经开发了TILLING作为一个服务于拟南芥社区被称为拟南芥TILLING项目（ATP）。我们的目标是根据国际研究界的要求，在目标1kb位点上快速提供乙基甲磺酸诱导突变的等位基因系列。在公开运作的第一年，ATP已经在拟南芥研究人员订购的>100个基因中发现、测序并传递了>1000个突变。这里描述的工具和方法可以适用于为其他生物体创建类似的设施。
TILLING ((T) under bar argeting (I) under bar nduced (L) under bar ocal (L) under bar esions in (G) under bar enomes) is a general reverse-genetic strategy that provides an allelic series of induced point mutations in genes of interest. High-throughput TILLING allows the rapid and low-cost discovery of induced point mutations in populations of chemically mutagenized individuals. As chemical mutagenesis is widely applicable and mutation detection for TILLING is dependent only on sufficient yield of PCR products, TILLING can be applied to most organisms. We have developed TILLING as a service to the Arabidopsis community known as the Arabidopsis TILLING Project (ATP). Our goal is to rapidly deliver allelic series of ethylmethanesulfonate-induced mutations in target 1-kb loci requested by the international research community. In the first year of public operation, ATP has discovered, sequenced, and delivered >1000 mutations in >100 genes ordered by Arabidopsis researchers. The tools and methodologies described here can be adapted to create similar facilities for other organisms.