The use of microRNAs as reference genes for quantitative polymerase chain reaction in soybean
The use of microRNAs as reference genes for quantitative polymerase chain reaction in soybean
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DOI：
10.1016/j.ab.2010.07.020
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发表时间：
2010-11-15
影响因子：
2.9
通讯作者：
Margis, Rogerio
中科院分区：
文献类型：
--
作者：
Kulcheski, Franceli Rodrigues;Marcelino-Guimaraes, Francismar Correa;Margis, Rogerio
逆转录实时定量聚合酶链反应（RT-qPCR）是用于研究基因表达的稳健且广泛应用的技术。然而，为了正确分析和解释结果，选择合适的基因用作内部对照是一个关键因素。这些基因，如管家基因，在不同组织和不同条件下应具有恒定的表达水平。基因组测序的进展提供了高通量的基因表达分析，并有助于鉴定新的基因，包括microRNA（miRNA）。miRNAs是真核生物基因组的基本调控基因，具有多种生物学功能。本研究对不同大豆组织和基因型以及非生物和生物胁迫处理后miRNA表达的稳定性进行了研究。本研究代表了第一次调查的适合miRNAs作为管家基因在植物中。将10种miRNAs的转录稳定性与先前报道的6种大豆管家基因的转录稳定性进行比较。在这项研究中，我们提供的证据表明，miR 156 b和miR 1520 d的表达稳定性是最高的大豆实验。此外，这些miRNAs基因比涉及RT-qPCR的大豆基因表达研究中最常用的蛋白质编码基因更稳定。(C)2010年爱思唯尔公司All rights reserved.
Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is a robust and widely applied technique used to investigate gene expression. However, for correct analysis and interpretation of results, the choice of a suitable gene to use as an internal control is a crucial factor. These genes, such as housekeeping genes, should have a constant expression level in different tissues and across different conditions. The advances in genome sequencing have provided high-throughput gene expression analysis and have contributed to the identification of new genes, including microRNAs (miRNAs). The miRNAs are fundamental regulatory genes of eukaryotic genomes, acting on several biological functions. In this study, miRNA expression stability was investigated in different soybean tissues and genotypes as well as after abiotic or biotic stress treatments. The present study represents the first investigation into the suitability of miRNAs as housekeeping genes in plants. The transcript stability of 10 miRNAs was compared to those of six previously reported housekeeping genes for the soybean. In this study, we provide evidence that the expression stabilities of miR156b and miR1520d were the highest across the soybean experiments. Furthermore, these miRNAs genes were more stable than the most commonly protein-coding genes used in soybean gene expression studies involving RT-qPCR. (C) 2010 Elsevier Inc. All rights reserved.