Komatsu Setsuko
University of Tsukuba, Tsukuba, 305-8572, Japan.
Methods Mol Biol. 2018;1696:81-90. doi: 10.1007/978-1-4939-7411-5_5.
The integrity of a subcellular proteomics is largely dependent on purity of the isolated compartment away from other contaminants. If high-purity nuclei is isolated, nuclear proteomics is a useful approach for investigating the mechanisms underlying plant physiological function. Although the isolation of high-purity nuclei from tissue or organ in plant is a difficult task, successful purification has been achieved through fractionation processes. For purification, there are five protocols such as (1) differential centrifugation, (2) discontinuous Percoll gradients, (3) continuous sucrose gradients, (4) combined continuous Percoll/sucrose gradients, and (5) continuous Percoll gradients. Furthermore, because purity assessment of purified nuclei is an important step, it is also described in this chapter.
亚细胞蛋白质组学的完整性在很大程度上取决于所分离的区室相对于其他污染物的纯度。如果分离出高纯度的细胞核，细胞核蛋白质组学就是研究植物生理功能潜在机制的一种有用方法。尽管从植物组织或器官中分离高纯度的细胞核是一项艰巨的任务，但通过分级分离过程已成功实现了纯化。为了进行纯化，有五种方案，例如（1）差速离心，（2）不连续的Percoll梯度，（3）连续的蔗糖梯度，（4）连续的Percoll/蔗糖梯度组合，以及（5）连续的Percoll梯度。此外，由于纯化细胞核的纯度评估是重要的一步，本章也对此进行了描述。
