Muskett Paul R, Clissold Leah, Marocco Adriano, Springer Patricia S, Martienssen Robert, Dean Caroline
Department of Cell and Developmental Biology, John Innes Centre, Colney, Norwich NR4 7UH, United Kingdom.
Plant Physiol. 2003 Jun;132(2):506-16. doi: 10.1104/pp.102.016535. Epub 2003 May 15.
We describe a new resource for targeted insertional mutagenesis in Arabidopsis using a maize (Zea mays) Activator/Dissociation (Ds) two-element system. The two components of the system, T-DNA vectors carrying a Ds launch pad and a stable Activator transposase source, were designed to simplify selection of transposition events and maximize their usefulness. Because Ds elements preferentially transpose to nearby genomic sites, they can be used in targeted mutagenesis of linked genes. To efficiently target all genes throughout the genome, we generated a large population of transgenic Arabidopsis plants containing the Ds launch pad construct, identified lines containing single Ds launch pad inserts, and mapped the positions of Ds launch pads in 89 lines. The integration sites of the Ds launch pads were relatively evenly distributed on all five chromosomes, except for a region of chromosomes 2 and 4 and the centromeric regions. This resource therefore provides access to the majority of the Arabidopsis genome for targeted tagging.
我们描述了一种利用玉米（Zea mays）激活子/解离（Ds）双元件系统在拟南芥中进行靶向插入诱变的新资源。该系统的两个组件，即携带Ds发射平台的T-DNA载体和稳定的激活子转座酶来源，旨在简化转座事件的筛选并最大限度地提高其效用。由于Ds元件优先转座到附近的基因组位点，它们可用于连锁基因的靶向诱变。为了有效地靶向整个基因组中的所有基因，我们生成了大量含有Ds发射平台构建体的转基因拟南芥植株，鉴定出含有单个Ds发射平台插入片段的株系，并绘制了89个株系中Ds发射平台的位置。Ds发射平台的整合位点在所有五条染色体上相对均匀分布，但2号和4号染色体的一个区域以及着丝粒区域除外。因此，该资源为靶向标签提供了访问拟南芥基因组大部分区域的途径。
