Validation of internal control genes for quantitative gene expression studies in chickpea (Cicer arietinum L.)
Validation of internal control genes for quantitative gene expression studies in chickpea (Cicer arietinum L.)
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DOI：
10.1016/j.bbrc.2010.04.079
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发表时间：
2010-05-28
影响因子：
3.1
通讯作者：
Jain, Mukesh
中科院分区：
文献类型：
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作者：
Garg, Rohini;Sahoo, Annapurna;Jain, Mukesh
实时聚合酶链反应（PCR）数据需要在所有实验条件下用恒定水平表达的内部对照基因进行标准化，以获得准确可靠的基因表达结果。本研究以鹰嘴豆（Cicer arietinum L.）为材料，对12个候选内参基因ACT 1、EF 1 α、GAPDH、IF 4a、TUB 6、UBC、UBQ 5、UBQ 10、18 SrRNA、25 SrRNA、GRX和HSP 90在不同器官、发育阶段和胁迫条件下的表达进行了研究。已经过验证。它们的表达水平在分析的各种组织样品中变化很大。EF 1 α和HSP 90的表达水平在分析的各种器官/发育阶段中最恒定。类似地，IF 4a和GAPDH的表达水平在各种胁迫条件下最恒定。一组两个最稳定的基因被发现足以在给定的一组鹰嘴豆组织样品的实时PCR数据的准确和可靠的标准化。在这项研究中确定的最恒定表达的基因应该是有用的鹰嘴豆中的各种组织样本的基因表达数据的归一化。(C)2010年爱思唯尔公司All rights reserved.
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, EF1 alpha, 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 EF1 alpha 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. (C) 2010 Elsevier Inc. All rights reserved.