A simple method for isolating import-competent Arabidopsis chloroplasts
A simple method for isolating import-competent Arabidopsis chloroplasts
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DOI：
10.1016/s0014-5793(02)03342-2
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发表时间：
2002-10-09
期刊：
影响因子：
3.5
通讯作者：
Jarvis, P
中科院分区：
文献类型：
--
作者：
Aronsson, H;Jarvis, P
我们提出了一种简单、快速、低成本的分离方法，可用于研究叶绿体蛋白质的输入。叶绿体分离的效率取决于植物组织的量与缓冲液体积之间的比率、均质化设备的大小和速度以及均质化烧杯的大小。进口方法证明是有用的，当表征不同的前体蛋白，发育阶段和进口缺陷突变体。通过时间进程实验，可以在线性范围内测量进口率。与原生质体相比，这种分离方法具有显著的时间和成本节省（分别约为80%和95%），并产生具有更高的蛋白质输入能力的叶绿体。(C)2002年欧洲生物化学学会联合会。由Elsevier Science B.V.出版，版权所有。
We present a simple, rapid and low-cost method for isolating a high yield of Arahidopsis chloroplasts that can be used to study chloroplast protein import. Efficiency of chloroplast isolation was dependent upon the ratio between amount of plant tissue and the buffer volume, the size and speed of the homogenisation equipment, and the size of the homogenisation beaker. The import method proved useful when characterising different precursor proteins, developmental stages and import-defective mutants. Time-course experiments enabled the measurement of import rates in the linear range. Compared to protoplastation, this isolation method has significant time and cost savings (similar to80% and similar to95%, respectively), and yields chloroplasts with a higher capacity to import proteins. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.