Chang Chao-Yan, Gou Jin-Ying
School of Life Sciences, Fudan University, Shanghai 200438, China.
College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
MethodsX. 2023 Jun 20;11:102266. doi: 10.1016/j.mex.2023.102266. eCollection 2023 Dec.
Thylakoids host a large number of proteins to confer photosynthesis and chemical biosynthesis essential for plant survival and growth. Successful isolation of high-quality thylakoids is the first step to studying the compositions and function of thylakoid protein and metabolites. Nevertheless, former studies isolated chloroplasts and thylakoids using a high-speed centrifuge with Percoll, which was expensive and unfriendly to the environment. The method presented here aims to establish a simple and inexpensive method to isolate high-quality thylakoids for protein analysis by utilizing sucrose instead of Percoll to reduce the cost and modify the centrifuge speed into the range usually used in labs.
类囊体含有大量蛋白质,这些蛋白质赋予植物生存和生长所必需的光合作用和化学合成作用。成功分离高质量的类囊体是研究类囊体蛋白质和代谢产物的组成及功能的第一步。然而,以前的研究使用配备有聚蔗糖的高速离心机来分离叶绿体和类囊体,这种方法既昂贵又不环保。本文提出的方法旨在建立一种简单且低成本的方法,通过使用蔗糖而非聚蔗糖来降低成本,并将离心机速度调整到实验室常用范围,从而分离高质量的类囊体用于蛋白质分析。