Selection of housekeeping genes for normalization by real-time RT-PCR:: Analysis of Or-MYB1 gene expression in Orobanche ramosa development
Selection of housekeeping genes for normalization by real-time RT-PCR:: Analysis of Or-MYB1 gene expression in Orobanche ramosa development
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DOI：
10.1016/j.ab.2008.05.003
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发表时间：
2008-08-15
影响因子：
2.9
通讯作者：
Roman, B.
中科院分区：
文献类型：
--
作者：
Gonzalez-Verdejo, C. I.;Die, J. V.;Roman, B.
实时荧光PCR已成为候选基因准确和深入表达研究的首选方法。为了避免偏倚，实时PCR是指一个或几个内部控制基因，不应该在治疗之间波动。寄生植物列当中参考基因的需求已经出现，在这方面的研究尚未进行评估。在这项研究中，使用BestKeeper软件程序比较了基因18 S rRNA、Or-act 1、Or-tub 1和Or-ubq 1的表达稳定性。在4个常见的内源控制基因中，Or-act 1和Or-ubq 1在O.分枝样本。与此同时，进行了一项研究，研究转录因子Or-MYB 1的表达，这似乎是在感染前阶段牵连。这里提出的标准化策略是准确的实时PCR表达谱的先决条件，除其他外，开辟了研究低拷贝数样转录因子的信使RNA水平的可能性。(C)2008年爱思唯尔公司All rights reserved.
Real-time PCR has become the method of choice for accurate and in-depth expression studies of candidate genes. To avoid bias, real-time PCR is referred to one or several internal control genes that should not fluctuate among treatments. A need for reference genes in the parasitic plant Orobanche ramosa has emerged, and the studies in this area have not yet been evaluated. In this study, the genes 18S rRNA, Or-act1, Or-tub1, and Or-ubq1 were compared in terms of expression stability using the BestKeeper software program. Among the four common endogenous control genes, Or-act1 and Or-ubq1 were the most stable in O. ramosa samples. In parallel, a study was carried out studying the expression of the transcription factor Or-MYB1 that seemed to be implicated during preinfection stages. The normalization strategy presented here is a prerequisite to accurate real-time PCR expression profiling that, among other things, opens up the possibility of studying messenger RNA levels of low-copy-number-like transcription factors. (C) 2008 Elsevier Inc. All rights reserved.