Department of Genetics, Development, and Cell Biology, Iowa State University, Ames, Iowa 50011, USA.
Plant Cell. 2010 Jun;22(6):1667-85. doi: 10.1105/tpc.109.073452. Epub 2010 Jun 25.
The maize (Zea mays) transposable element Dissociation (Ds) was mobilized for large-scale genome mutagenesis and to study its endogenous biology. Starting from a single donor locus on chromosome 10, over 1500 elements were distributed throughout the genome and positioned on the maize physical map. Genetic strategies to enrich for both local and unlinked insertions were used to distribute Ds insertions. Global, regional, and local insertion site trends were examined. We show that Ds transposed to both linked and unlinked sites and displayed a nonuniform distribution on the genetic map around the donor r1-sc:m3 locus. Comparison of Ds and Mutator insertions reveals distinct target preferences, which provide functional complementarity of the two elements for gene tagging in maize. In particular, Ds displays a stronger preference for insertions within exons and introns, whereas Mutator insertions are more enriched in promoters and 5'-untranslated regions. Ds has no strong target site consensus sequence, but we identified properties of the DNA molecule inherent to its local structure that may influence Ds target site selection. We discuss the utility of Ds for forward and reverse genetics in maize and provide evidence that genes within a 2- to 3-centimorgan region flanking Ds insertions will serve as optimal targets for regional mutagenesis.
玉米(Zea mays)转座因子解离(Ds)被用于大规模的基因组诱变和研究其内源生物学。从 10 号染色体上的一个单一供体位点开始,超过 1500 个元件分布在整个基因组中,并定位在玉米物理图谱上。为了丰富局部和非连锁插入,我们使用了遗传策略来分配 Ds 插入。检查了全局、区域和局部插入位点趋势。我们表明,Ds 转座到连锁和非连锁位点,并在供体 r1-sc:m3 基因座周围的遗传图谱上显示出非均匀的分布。Ds 和 Mutator 插入的比较揭示了不同的靶标偏好,这为玉米中的基因标记提供了这两个元件的功能互补性。特别是,Ds 显示出对内含子和外显子内插入的更强偏好,而 Mutator 插入在启动子和 5'非翻译区中更为丰富。Ds 没有强烈的靶标序列共识,但我们确定了 DNA 分子固有的局部结构特性,这些特性可能影响 Ds 靶标选择。我们讨论了 Ds 在玉米正向和反向遗传学中的应用,并提供了证据表明,Ds 插入侧翼 2-3 厘摩区域内的基因将作为区域诱变的最佳靶标。