A method for isolating a high yield of Arabidopsis chloroplasts capable of efficient import of precursor proteins
A method for isolating a high yield of Arabidopsis chloroplasts capable of efficient import of precursor proteins
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DOI：
10.1046/j.0960-7412.2001.01061.x
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发表时间：
2001-07-01
期刊：
影响因子：
7.2
通讯作者：
Keegstra, K
中科院分区：
文献类型：
--
作者：
Fitzpatrick, LM;Keegstra, K
在不同条件下，采用不同的方法，从拟南芥植株中分离出叶绿体，以确定一种能够结合和导入前体蛋白的叶绿体方法。从原生质体中分离出来的叶绿体经Percoll梯度纯化后具有很强的进口能力，前体的非特异性结合很少，完整叶绿体的产量很高（0.1 mg叶绿素/g FW）。在琼脂板上生长的植物的叶绿体比在土壤上生长的植物的叶绿体进口率高得多。在平板栽培植物叶绿体的所有年龄测试中，蛋白质进口量保持较高，而在土壤栽培植物的发育过程中，蛋白质进口量下降。拟南芥叶绿体输入一系列前体蛋白，并具有与豌豆叶绿体结合和输入的核苷酸需求。
Chloroplasts were isolated from Arabidopsis plants grown under different conditions, and using different protocols, to determine a method that would yield chloroplasts capable of binding and importing precursor proteins. Chloroplasts isolated from protoplasts and purified on a Percoll gradient were highly import-competent, with little non-specific binding of the precursor, and a high yield of intact chloroplasts (0.1 mg chlorophyll/g FW). Chloroplasts from plants grown on agar plates had a much higher rate of import than those from plants grown on soil. Protein import remained high at all of the ages tested for chloroplasts from plate-grown plants, whereas it declined during the development of soil-grown plants. Arabidopsis chloroplasts imported a range of precursor proteins and had nucleotide requirements for binding and import similar to those reported for pea chloroplasts.