National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi 110 067, India.
Biochem Biophys Res Commun. 2010 May 28;396(2):283-8. doi: 10.1016/j.bbrc.2010.04.079. Epub 2010 Apr 23.
The real-time polymerase chain reaction (PCR) data requires normalization with an internal control gene expressed at constant levels under all the experimental conditions being analyzed for accurate and reliable gene expression results. In this study, the expression of 12 candidate internal control genes, including ACT1, EF1alpha, GAPDH, IF4a, TUB6, UBC, UBQ5, UBQ10, 18SrRNA, 25SrRNA, GRX and HSP90, in a diverse set of 18 tissue samples representing different organs/developmental stages and stress conditions in chickpea (Cicer arietinum L.) has been validated. Their expression levels vary considerably in various tissue samples analyzed. The expression levels of EF1alpha and HSP90 are most constant across various organs/developmental stages analyzed. Similarly, the expression levels of IF4a and GAPDH are most constant across various stress conditions. A set of two most stable genes is found sufficient for accurate and reliable normalization of real-time PCR data in the given set of tissue samples of chickpea. The genes with most constant expression identified in this study should be useful for normalization of gene expression data in a wide variety of tissue samples in chickpea.
实时聚合酶链反应（PCR）数据需要用在分析的所有实验条件下均以恒定水平表达的内参基因进行标准化，以获得准确可靠的基因表达结果。在这项研究中，在 chickpea（鹰嘴豆）的 18 个不同组织样本中，对包括 ACT1、EF1alpha、GAPDH、IF4a、TUB6、UBC、UBQ5、UBQ10、18SrRNA、25SrRNA、GRX 和 HSP90 在内的 12 个候选内参基因的表达进行了验证，这些样本代表了不同的器官/发育阶段和胁迫条件。在分析的各种组织样本中，它们的表达水平差异很大。EF1alpha 和 HSP90 的表达水平在分析的各种器官/发育阶段中最稳定。同样，IF4a 和 GAPDH 的表达水平在各种胁迫条件下最稳定。在给定的 chickpea 组织样本集中，发现一组两个最稳定的基因足以准确可靠地对实时 PCR 数据进行标准化。本研究中确定的表达最稳定的基因对于 chickpea 中各种组织样本的基因表达数据的标准化应该是有用的。
