Vandenbussche Michiel, Zethof Jan, Gerats Tom
UMR 5667 CNRS-INRA-ENS Lyon-Unversité Lyon I, RDP Laboratory, ENS Lyon, Lyon, Cedex, France.
Methods Mol Biol. 2013;1057:239-50. doi: 10.1007/978-1-62703-568-2_17.
Transposon tagging has been used successfully in a range of organisms for the cloning of mutants of interest. In species containing high copy numbers of transposable elements combined with a high transposition rate, forward cloning can be quite challenging and requires specific high-resolution methods. Here we detail an updated version of the Transposon Display technique, which allows visualization of large numbers of transposon-flanking sequences simultaneously in a highly robust and reproducible manner. This strategy was developed for the analysis of the transpositional behavior of the dTph1 transposon and for the forward cloning of mutants, particularly in the Petunia W138 background, individuals of which can contain >200 copies of the endogenous dTph1 element. The method is derived from the AFLP technique and can in principle easily be adapted to any system.
转座子标签法已成功应用于一系列生物体中，用于克隆感兴趣的突变体。在含有高拷贝数转座元件且转座率高的物种中，正向克隆颇具挑战性，需要特定的高分辨率方法。在此，我们详细介绍转座子展示技术的更新版本，该技术能够以高度稳健且可重复的方式同时可视化大量转座子侧翼序列。此策略是为分析dTph1转座子的转座行为以及正向克隆突变体而开发的，特别是在矮牵牛W138背景中，该背景下的个体可能含有>200个拷贝的内源性dTph1元件。该方法源自AFLP技术，原则上可轻松适用于任何系统。
