多核糖体的纯化

Purification of polysomes.

作者信息

Rivera Maria C, Maguire Bruce, Lake James A

机构信息

Department of Biology and Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, Virginia 23284;

Primary Pharmacology Group, Pfizer Global Research and Development, Groton, Connecticut 06340;

出版信息

Cold Spring Harb Protoc. 2015 Mar 2;2015(3):303-5. doi: 10.1101/pdb.prot081364.

DOI:10.1101/pdb.prot081364

PMID:25734067

Abstract

Isolated polysomes (also known as translating ribosomes or polyribosomes) are mRNA-ribosome complexes that are frequently used for the in vitro study of the regulation of protein synthesis. Here we describe a protocol for the isolation of prokaryotic and plant polysomes by sucrose gradient sedimentation. The protocol allows for the separation of multiple ribosomes attached to mRNA from run-off ribosome monomers. In addition to collecting the polysome pellet, if the sucrose gradients used in this protocol are fractionated, it is possible to isolate and quantify the polysomes, the ribosome monomers, and the ribosomal subunits.

摘要

游离多核糖体（也称为正在翻译的核糖体或多聚核糖体）是mRNA-核糖体复合物，常用于蛋白质合成调控的体外研究。在此，我们描述了一种通过蔗糖梯度沉降分离原核生物和植物多核糖体的方法。该方法能够将附着在mRNA上的多个核糖体与游离核糖体单体分离。除了收集多核糖体沉淀外，如果对本方法中使用的蔗糖梯度进行分级分离，还可以分离并定量多核糖体、核糖体单体和核糖体亚基。