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利用exon-targeting 内源性逆转录转座子 LORE1 建立百脉根基因打靶群体。

Establishment of a Lotus japonicus gene tagging population using the exon-targeting endogenous retrotransposon LORE1.

机构信息

Division of Plant Sciences, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan.

出版信息

Plant J. 2012 Feb;69(4):720-30. doi: 10.1111/j.1365-313X.2011.04826.x. Epub 2011 Dec 12.

Abstract

We established a gene tagging population of the model legume Lotus japonicus using an endogenous long terminal repeat (LTR) retrotransposon Lotus Retrotransposon 1 (LORE1). The population was composed of 2450 plant lines, from which a total of 4532 flanking sequence tags of LORE1 were recovered by pyrosequencing. The two-dimensional arrangement of the plant population, together with the use of multiple identifier sequences in the primers used to amplify the flanking regions, made it possible to trace insertions back to the original plant lines. The large-scale detection of new LORE1 insertion sites revealed a preference for genic regions, especially in exons of protein-coding genes, which is an interesting feature to consider in the interaction between host genomes and chromoviruses, to which LORE1 belongs, a class of retrotransposon widely distributed among plants. Forward screening of the symbiotic mutants from the population succeeded to identify five symbiotic mutants of known genes. These data suggest that LORE1 is robust as a genetic tool.

摘要

我们利用内源性长末端重复(LTR)反转录转座子 Lotus Retrotransposon 1(LORE1)建立了模式豆科植物百脉根的基因标记群体。该群体由 2450 条植物品系组成,通过焦磷酸测序共回收了 4532 个 LORE1 的侧翼序列标签。植物群体的二维排列,以及在用于扩增侧翼区域的引物中使用多个标识符序列,使得能够将插入回溯到原始植物品系。大规模检测新的 LORE1 插入位点揭示了其对基因区域的偏好,特别是在蛋白质编码基因的外显子中,这是一个有趣的特征,考虑到宿主基因组和 Chromoviruses(LORE1 属于 Chromoviruses 类反转录转座子,广泛分布于植物中)之间的相互作用。对群体中的共生突变体进行正向筛选成功鉴定了五个已知基因的共生突变体。这些数据表明 LORE1 作为一种遗传工具具有很强的稳健性。

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