Samanta Subhasis, Thakur Jitendra Kumar
National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi, India.
Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, India.
Methods Mol Biol. 2017;1629:123-140. doi: 10.1007/978-1-4939-7125-1_9.
The Mediator complex is a multi-protein complex that acts as a molecular bridge conveying transcriptional messages from the cis element-bound transcription factor to the RNA Polymerase II machinery. It is found in all eukaryotes including members of the plant kingdom. Increasing number of reports from plants regarding different Mediator subunits involved in a multitude of processes spanning from plant development to environmental interactions have firmly established it as a central hub of plant regulatory networks. Routine isolation of Mediator complex in a particular species is a necessity because of many reasons. First, composition of the Mediator complex varies from species to species. Second, the composition of the Mediator complex in a particular species is not static under all developmental and environmental conditions. Besides this, at times, Mediator complex is used in in vitro transcription systems. Rice, a staple food crop of the world, is used as a model monocot crop. Realizing the need of a reliable protocol for the isolation of Mediator complex from plants, we describe here the isolation of Mediator complex from rice.
中介体复合物是一种多蛋白复合物，作为分子桥梁，将转录信息从与顺式元件结合的转录因子传递到RNA聚合酶II机制。它存在于所有真核生物中，包括植物界的成员。越来越多来自植物的关于不同中介体亚基参与从植物发育到环境相互作用等众多过程的报道，已将其牢固地确立为植物调控网络的中心枢纽。由于多种原因，在特定物种中常规分离中介体复合物是必要的。首先，中介体复合物的组成因物种而异。其次，特定物种中介体复合物的组成在所有发育和环境条件下并非一成不变。除此之外，有时中介体复合物会用于体外转录系统。水稻是世界主要粮食作物，用作单子叶植物的模式作物。意识到从植物中分离中介体复合物需要可靠方案，我们在此描述从水稻中分离中介体复合物的方法。
