National Institute for Basic Biology, Nishigonaka 38, Myodaiji, Okazaki, Aichi, 444-8585, Japan.
Mol Genet Genomics. 2010 Nov;284(5):343-55. doi: 10.1007/s00438-010-0569-9. Epub 2010 Sep 10.
The nonautonomous nDart1 element in the hAT superfamily is one of a few active DNA transposons in rice. Its transposition can be induced by crossing with a line containing an active autonomous element, aDart1, and stabilized by segregating aDart1. No somaclonal variation should occur in nDart1-promoted gene tagging because no tissue culture is involved in nDart1 activation. By transposon display analysis, we examined the activities of nDart1-related elements in the selfed progeny of a mutable virescent pyl-v plant containing aDart1. Although various nDart1-related elements are present in the rice genome, only nDart1-3 subgroup elements, nDart1-0 and nDart1-3 in particular, were found to be transposed frequently and integrated into various sites almost all over the genome, and a fraction of the transposed elements were found to be transmitted to the next generation. More than half of the newly integrated elements were identified as nDart1-0. Analysis of the newly inserted sites revealed that the nDart1-3 subgroup elements were predominantly integrated into single-copy regions. More than 60% of the transposed elements were inserted into the genic regions that comprise putative coding regions and their 0.5-kb flanking segments, and approximately two-thirds of them were within the 0.5-kb area in front of the putative initiation codons, i.e., promoter-proximal genic regions. These characteristic features of nDart1-3 subgroup elements seem to be suitable for developing an efficient and somaclonal variation-free gene tagging system for rice functional genomics.
hAT 超家族中的非自主 nDart1 元件是水稻中少数几种活跃的 DNA 转座子之一。它的转座可以通过与含有活性自主元件 aDart1 的系杂交来诱导,并通过分离 aDart1 来稳定。在 nDart1 促进的基因标记中不应发生体细胞变异,因为在 nDart1 激活中不涉及组织培养。通过转座子展示分析,我们检查了含有 aDart1 的可变性绿黄质 pyl-v 植物的自交后代中 nDart1 相关元件的活性。尽管各种 nDart1 相关元件存在于水稻基因组中,但只有 nDart1-3 亚组元件,特别是 nDart1-0 和 nDart1-3,被发现频繁转座并整合到基因组的各种位点,并且一部分转座元件被传递到下一代。新整合的元件中有一半以上被鉴定为 nDart1-0。新插入位点的分析表明,nDart1-3 亚组元件主要整合到单拷贝区域。超过 60%的转座元件插入到包含假定编码区及其 0.5kb 侧翼片段的基因区域,其中大约三分之二位于假定起始密码子前面的 0.5kb 区域内,即启动子近端基因区域。nDart1-3 亚组元件的这些特征似乎适合开发用于水稻功能基因组学的高效和无体细胞变异的基因标记系统。