Bock R
Institut für Biologie III, Universität Freiburg, Germany.
Methods. 1998 May;15(1):75-83. doi: 10.1006/meth.1998.0607.
Chloroplast mRNAs can be subject to posttranscriptional pyrimidine-to-pyrimidine conversions at highly specific sites. This RNA modification mechanism shows a high degree of similarity to plant mitochondrial editing but differs markedly from, and is most likely evolutionarily unrelated to, all other RNA editing systems. The study of RNA editing processes in chloroplasts has been largely hampered by the lack of in vitro editing systems; however, considerable insights into the recognition mechanisms of individual editing sites have come from in vivo approaches. Chloroplast transformation proved to be a particularly useful tool to study plastid RNA editing. In this article, specific methods for the analysis of chloroplast RNA editing are discussed. Detailed experimental procedures are provided for (i) the purification of chloroplasts and (ii) the stable genetic transformation of higher plant plastids.
叶绿体mRNA在高度特异的位点可发生转录后嘧啶到嘧啶的转换。这种RNA修饰机制与植物线粒体编辑表现出高度相似性，但与所有其他RNA编辑系统明显不同，并且很可能在进化上没有关联。叶绿体中RNA编辑过程的研究在很大程度上受到缺乏体外编辑系统的阻碍；然而，通过体内方法已对个别编辑位点的识别机制有了相当深入的了解。叶绿体转化被证明是研究质体RNA编辑的一种特别有用的工具。本文讨论了分析叶绿体RNA编辑的具体方法。提供了用于（i）叶绿体纯化和（ii）高等植物质体稳定遗传转化的详细实验步骤。
