Kempken Frank, Bolle Nina, Forner Joachim, Binder Stefan
Genetische Botanik und Molekularbiologie, Botanisches Institut und Botanischer Garten, Christian-Albrechts-Universität, Kiel, Germany.
Methods Mol Biol. 2007;372:177-92. doi: 10.1007/978-1-59745-365-3_13.
Mitochondria are genetic compartments with their own enzymatic equipment for maintenance and expression of their genetic information. As in all genetic systems, gene expression has to be regulated, and in mitochondria this also has to be coordinated with the expression of nuclear-encoded mitochondrial proteins. Presently, there is virtually no information available about the mechanistic details and the enzymes involved in these processes. There is still much to be learned about how plant mitochondrial gene expression is managed and to what extent the contribution of transcription initiation and posttranscriptional processes, respectively, contribute to this control. As one prerequisite for better understanding of the mechanisms and regulatory controls, more fundamental data on mitochondrial transcription initiation and posttranscriptional RNA processing are necessary. As part of the essential methodology, we present methods for the analysis of the 5' and 3' extremities of mitochondrial transcripts and the identification of transcription initiation sites. An in organello system is described for the functional investigation of ribonucleic acid editing in plant mitochondria.
线粒体是具有自身用于维持和表达遗传信息的酶设备的遗传区室。与所有遗传系统一样，基因表达必须受到调控，而在线粒体中，这还必须与核编码的线粒体蛋白的表达相协调。目前，关于这些过程的机制细节和所涉及的酶几乎没有可用信息。关于植物线粒体基因表达是如何管理的，以及转录起始和转录后过程分别在多大程度上对这种控制有贡献，仍有许多有待了解的地方。作为更好地理解机制和调控控制的一个先决条件，关于线粒体转录起始和转录后RNA加工的更基础数据是必要的。作为基本方法的一部分，我们介绍了分析线粒体转录本5'和3'末端以及鉴定转录起始位点的方法。描述了一种用于植物线粒体中核糖核酸编辑功能研究的体外系统。
