Aronsson Henrik, Jarvis Paul
Department of Biology, University of Leicester, University Road, LE1 7RH, Leicester, UK
FEBS Lett. 2002 Oct 9;529(2-3):215-20. doi: 10.1016/s0014-5793(02)03342-2.
We present a simple, rapid and low-cost method for isolating a high yield of Arabidopsis 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 (approximately 80% and approximately 95%, respectively), and yields chloroplasts with a higher capacity to import proteins.
我们提出了一种简单、快速且低成本的方法来分离高产拟南芥叶绿体，该方法可用于研究叶绿体蛋白输入。叶绿体分离效率取决于植物组织量与缓冲液体积之比、匀浆设备的大小和速度以及匀浆烧杯的大小。当对不同的前体蛋白、发育阶段和输入缺陷型突变体进行表征时，该输入方法被证明是有用的。时间进程实验能够测量线性范围内的输入速率。与原生质体形成相比，这种分离方法在时间和成本上有显著节省（分别约为80%和约95%），并且产生的叶绿体具有更高的蛋白输入能力。
