A collection of 11,800 single-copy Ds transposon insertion lines in Arabidopsis
A collection of 11,800 single-copy Ds transposon insertion lines in Arabidopsis
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DOI：
10.1111/j.1365.313x.2004.02009.x
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发表时间：
2004-03-01
期刊：
影响因子：
7.2
通讯作者：
Shinozaki, K
中科院分区：
文献类型：
--
作者：
Kuromori, T;Hirayama, T;Shinozaki, K
利用Activator（Ac）/Dissociation（Ds）系统构建了10 000多个转座子标记株系，旨在收集拟南芥的插入突变体，为拟南芥功能基因组学研究提供有用的资源。使用半自动化方法通过高通量分析确定了11 800个独立株系中Ds元件的侧翼序列。序列数据使我们能够在每个品系中定位拟南芥基因组上的独特插入位点。7566个株系的Ds元件被插入到编码区或其附近，可能影响25000个拟南芥基因中的5031个基因的功能。一半的品系在1号染色体（Chr. 1）上具有Ds插入，其中供体品系具有供体位点。在另一半中，Ds插入分布在其他四条染色体上。Ds插入的染色体内分布随供体系的不同而不同。我们发现在每条染色体的末端附近都有Ds转座的热点，并且我们发现在核苷酸水平上对Ds插入靶点有一些统计学偏好。在对11800株系Ds插入位点进行系统分析的基础上，提出了利用Ds标记的单插入系进行功能基因组表型系统分析的设想。我们已经建立了插入位点序列和突变基因的可检索数据库（http：//rarge.gsc.riken.go.jp/），并将这些品系作为可用资源保存在RIKEN生物资源中心（http：//www.brc.riken.go.jp/Eng/）。
More than 10 000 transposon-tagged lines were constructed by using the Activator (Ac)/Dissociation (Ds) system in order to collect insertional mutants as a useful resource for functional genomics of Arabidopsis. The flanking sequences of the Ds element in the 11 800 independent lines were determined by high-throughput analysis using a semi-automated method. The sequence data allowed us to map the unique insertion site on the Arabidopsis genome in each line. The Ds element of 7566 lines is inserted in or close to coding regions, potentially affecting the function of 5031 of 25 000 Arabidopsis genes. Half of the lines have Ds insertions on chromosome 1 (Chr. 1), in which donor lines have a donor site. In the other half, the Ds insertions are distributed throughout the other four chromosomes. The intrachromosomal distribution of Ds insertions varies with the donor lines. We found that there are hot spots for Ds transposition near the ends of every chromosome, and we found some statistical preference for Ds insertion targets at the nucleotide level. On the basis of systematic analysis of the Ds insertion sites in the 11 800 lines, we propose the use of Ds-tagged lines with a single insertion in annotated genes for systematic analysis of phenotypes (phenome analysis) in functional genomics. We have opened a searchable database of the insertion-site sequences and mutated genes (http://rarge.gsc.riken.go.jp/) and are depositing these lines in the RIKEN BioResource Center as available resources (http://www.brc.riken.go.jp/Eng/).