Tirichine Leïla, Lin Xin, Thomas Yann, Lombard Bérangère, Loew Damarys, Bowler Chris
Environmental and Evolutionary Genomics Section, Institut de Biologie de l'École Normale Supérieure (IBENS), CNRS UMR 8197 INSERM U1024, 46 rue d'Ulm 75005 Paris, France.
Environmental and Evolutionary Genomics Section, Institut de Biologie de l'École Normale Supérieure (IBENS), CNRS UMR 8197 INSERM U1024, 46 rue d'Ulm 75005 Paris, France.
Mar Genomics. 2014 Feb;13:21-5. doi: 10.1016/j.margen.2013.11.006. Epub 2013 Dec 4.
Post-translational modifications of histones affect many biological processes by influencing higher order chromatin structure that affects gene and genome regulation. It is therefore important to develop methods for extracting histones while maintaining their native post-translational modifications. While histone extraction protocols have been developed in multicellular and single celled organisms such as yeast and Arabidopsis, they are inefficient in diatoms that have a silica cell wall that is likely to hinder histone extraction. We report in this work a rapid and reliable method for extraction of large amounts of high quality histones from the two model diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. The protocol is an important enabling step permitting downstream applications such as western blotting and mass spectrometry.
组蛋白的翻译后修饰通过影响高级染色质结构来影响许多生物学过程，而高级染色质结构又会影响基因和基因组调控。因此，开发在维持组蛋白天然翻译后修饰的同时提取组蛋白的方法非常重要。虽然已经在多细胞和单细胞生物（如酵母和拟南芥）中开发了组蛋白提取方案，但在具有可能阻碍组蛋白提取的硅质细胞壁的硅藻中，这些方法效率不高。我们在这项工作中报告了一种从两种模式硅藻三角褐指藻和假微型海链藻中快速可靠地提取大量高质量组蛋白的方法。该方案是一个重要的起始步骤，可用于蛋白质免疫印迹和质谱分析等下游应用。
