Kleine Tatjana, Leister Dario
Lehrstuhl für Molekularbiologie der Pflanzen (Botanik), Department Biologie I, Ludwig-Maximilians-Universität, Planegg-Martinsried, Munich, Germany.
Methods Mol Biol. 2011;775:117-34. doi: 10.1007/978-1-61779-237-3_7.
The vast majority of the several thousands of chloroplast proteins are encoded by nuclear genes. Regulation of their expression involves control of their transcription, and thus requires the transmission of information from chloroplast to nucleus (retrograde signalling). The most powerful approach to the analysis of the transcriptional regulation of chloroplast functions involves RNA hybridization to microarrays representing almost all nuclear genes of Arabidopsis thaliana, followed by statistical data analysis. This chapter provides detailed protocols for the preparation of RNA for microarray experiments, in particular the widely used Affymetrix ATH1 array. Finally, the use of the publicly available program Robin for statistical data analysis, as well as approaches to confirm microarray data, is introduced.
数千种叶绿体蛋白中的绝大多数是由核基因编码的。它们表达的调控涉及转录控制，因此需要从叶绿体向细胞核传递信息（逆向信号传导）。分析叶绿体功能转录调控的最有效方法是将RNA与代表拟南芥几乎所有核基因的微阵列进行杂交，然后进行统计数据分析。本章提供了用于微阵列实验的RNA制备的详细方案，特别是广泛使用的Affymetrix ATH1阵列。最后，介绍了使用公开可用的Robin程序进行统计数据分析以及确认微阵列数据的方法。
