Alba Rob, Fei Zhangjun, Payton Paxton, Liu Yang, Moore Shanna L, Debbie Paul, Cohn Jonathan, D'Ascenzo Mark, Gordon Jeffrey S, Rose Jocelyn K C, Martin Gregory, Tanksley Steven D, Bouzayen Mondher, Jahn Molly M, Giovannoni Jim
Boyce Thompson Institute for Plant Research, Cornell University Campus, Ithaca, NY, USA.
Plant J. 2004 Sep;39(5):697-714. doi: 10.1111/j.1365-313X.2004.02178.x.
Gene expression profiling holds tremendous promise for dissecting the regulatory mechanisms and transcriptional networks that underlie biological processes. Here we provide details of approaches used by others and ourselves for gene expression profiling in plants with emphasis on cDNA microarrays and discussion of both experimental design and downstream analysis. We focus on methods and techniques emphasizing fabrication of cDNA microarrays, fluorescent labeling, cDNA hybridization, experimental design, and data processing. We include specific examples that demonstrate how this technology can be used to further our understanding of plant physiology and development (specifically fruit development and ripening) and for comparative genomics by comparing transcriptome activity in tomato and pepper fruit.
基因表达谱分析在剖析生物过程背后的调控机制和转录网络方面具有巨大潜力。本文我们详细介绍了他人和我们自己在植物基因表达谱分析中所使用的方法，重点是cDNA微阵列，并讨论了实验设计和下游分析。我们专注于强调cDNA微阵列制备、荧光标记、cDNA杂交、实验设计和数据处理的方法和技术。我们给出了具体实例，展示了该技术如何用于增进我们对植物生理学和发育（特别是果实发育和成熟）的理解，以及如何通过比较番茄和辣椒果实中的转录组活性用于比较基因组学研究。
