Monitoring global messenger RNA changes in externally controlled microarray experiments
Monitoring global messenger RNA changes in externally controlled microarray experiments
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DOI：
10.1038/sj.embor.embor798
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发表时间：
2003-04-01
期刊：
影响因子：
7.7
通讯作者：
Holstege, FCP
中科院分区：
文献类型：
--
作者：
van de Peppel, J;Kemmeren, P;Holstege, FCP
表达谱分析是一种通用的工具，具有一系列受益于差异基因表达的准确测定的应用。为了允许使用内源性转录物水平进行标准化，当前的微阵列分析假设相对较少的转录物变化，或者发生的任何变化都是平衡的。当使用内源基因进行标准化时，相对于大多数转录物的行为计算表达水平的变化。如果信使RNA群体发生全球变化，这并不反映绝对变化。使用外部RNA对照，我们建立了微阵列实验来监测全球变化。当包括外部对照时，发现大多数mRNA的水平在酵母稳定期和人热休克期间发生变化。即使是很小的全局变化也会对报告为差异表达的基因数量产生重大影响。这表明，全球mRNA的变化发生更频繁，比目前假设的，并表明，监测这种影响可能是重要的准确测定基因表达的变化。
Expression profiling is a universal tool, with a range of applications that benefit from the accurate determination of differential gene expression. To allow normalization using endogenous transcript levels, current microarray analyses assume that relatively few transcripts vary, or that any changes that occur are balanced. When normalization using endogenous genes is carried out, changes in expression levels are calculated relative to the behaviour of most of the transcripts. This does not reflect absolute changes if global shifts in messenger RNA populations occur. Using external RNA controls, we have set up microarray experiments to monitor global changes. The levels of most mRNAs were found to change during yeast stationary phase and human heat shock when external controls were included. Even small global changes had a significant effect on the number of genes reported as being differentially expressed. This suggests that global mRNA changes occur more frequently than is assumed at present, and shows that monitoring such effects may be important for the accurate determination of changes in gene expression.