Domanski Michal, LaCava John
Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland.
Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, USA.
Bio Protoc. 2017 Apr 20;7(8). doi: 10.21769/BioProtoc.2238.
The RNA exosome complex plays a central role in RNA processing and regulated turnover. Present both in cytoplasm and nucleus, the exosome functions through associations with ribonucleases and various adapter proteins (reviewed in [Kilchert ., 2016]). The following protocol describes an approach to purify RNA exosome complexes from HEK-293 cells, making use of inducible ectopic expression, affinity capture, and rate-zonal centrifugation. The obtained RNA exosomes have been used successfully for proteomic, structural, and enzymatic studies (Domanski ., 2016).
RNA外切体复合物在RNA加工和调控周转中起核心作用。外切体存在于细胞质和细胞核中，通过与核糖核酸酶和各种衔接蛋白结合发挥作用（见[Kilchert ., 2016]综述）。以下方案描述了一种从HEK-293细胞中纯化RNA外切体复合物的方法，该方法利用了诱导性异位表达、亲和捕获和速率区带离心。所获得的RNA外切体已成功用于蛋白质组学、结构和酶学研究（Domanski ., 2016）。
