Institut de Biologie de l'Ecole Normale Supérieure (IBENS), Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM), Ecole Normale Supérieure, PSL Research University, Paris, France.
Methods Mol Biol. 2021;2250:141-155. doi: 10.1007/978-1-0716-1134-0_14.
Transposable elements (TEs) are repetitive DNA sequences that have the ability to mobilize in the genome and create major effect mutations. Despite the importance of transposition as a source of genetic novelty, we still know little about the rate, landscape, and consequences of TE mobilization. This situation stems in large part from the repetitive nature of TEs, which complicates their analysis. Moreover, TE mobilization is typically rare and therefore new TE (i.e., non-reference) insertions tend to be missed in small-scale population studies. This chapter describes a TE-sequence capture approach designed to identify transposition events for most of the TE families that are potentially active in Arabidopsis thaliana. We show that our TE-sequence capture design provides an efficient means to detect with high sensitivity and specificity insertions that are present at a frequency as low as 1/1000 within a DNA sample.
转座元件 (TEs) 是具有在基因组中移动并产生重大效应突变能力的重复 DNA 序列。尽管转座作为遗传新颖性的来源非常重要,但我们对 TE 移动的速率、景观和后果仍然知之甚少。这种情况在很大程度上源于 TEs 的重复性质,这使得它们的分析变得复杂。此外,TE 移动通常很少见,因此在小规模的群体研究中,通常会错过新的 TE(即非参考)插入。本章描述了一种 TE 序列捕获方法,旨在鉴定拟南芥中可能活跃的大多数 TE 家族的转座事件。我们表明,我们的 TE 序列捕获设计提供了一种有效的手段,可以以高灵敏度和特异性检测 DNA 样本中频率低至 1/1000 的插入。