Chelysheva Liudmila A, Grandont Laurie, Grelon Mathilde
Institut National de la Recherche Agronomique (INRA), Centre de Versailles-Grignon, Versailles Cedex, France.
Methods Mol Biol. 2013;990:93-101. doi: 10.1007/978-1-62703-333-6_9.
Plant meiosis studies have enjoyed a fantastic boom in recent years with the use of Arabidopsis thaliana as an important model species for developmental studies because of its small genome, short life cycle, and large mutant collections. Unlike other eukaryotic models, plant meiosis does not display strict checkpoints and rarely commits to apoptotic processes, which makes it possible to investigate the whole meiotic process (spanning from premeiotic interphase to spore formation) in knockout mutants. In this chapter we describe a protocol for immunolabelling Arabidopsis and Brassica meiotic proteins on robustly spread chromosomes. This protocol allows the detection of a large range of proteins on well-preserved chromosomes and throughout the entire meiotic process.
近年来,由于拟南芥基因组小、生命周期短且有大量突变体库,作为发育研究的重要模式物种,植物减数分裂研究蓬勃发展。与其他真核生物模式不同,植物减数分裂不显示严格的检查点,很少进入凋亡过程,这使得在敲除突变体中研究整个减数分裂过程(从减数分裂前的间期到孢子形成)成为可能。在本章中,我们描述了一种在充分铺展的染色体上对拟南芥和油菜减数分裂蛋白进行免疫标记的方法。该方法能够在保存良好的染色体上以及整个减数分裂过程中检测到多种蛋白质。