Lecampion Cécile, Lanet Elodie, Robaglia Christophe
Laboratoire de Génétique et Biophysique des Plantes, Aix Marseille Univ, Marseille, F-13009, France.
CNRS, UMR 7265 Biologie Végétale et Microbiologie Environnementale, Marseille, F-13009, France.
Methods Mol Biol. 2017;1640:137-143. doi: 10.1007/978-1-4939-7165-7_9.
Argonaute (AGO) proteins play a key role in RNA silencing mechanisms. RNA silencing affects both RNA degradation and translation. The characterization of translation-associated RNA silencing mechanisms and components often requires polysome isolation and analysis. In this chapter, we describe the identification of AGO1 association with polysomes through polysome fractionation on sucrose gradient, preparation of proteins by filtration and concentration, and immunoblotting.
AGO蛋白在RNA沉默机制中起关键作用。RNA沉默影响RNA降解和翻译。对与翻译相关的RNA沉默机制和成分的表征通常需要进行多核糖体分离和分析。在本章中，我们描述了通过蔗糖梯度多核糖体分级分离、过滤和浓缩制备蛋白质以及免疫印迹来鉴定AGO1与多核糖体的关联。
