Reference gene selection for quantitative gene expression studies during biological invasions: A test on multiple genes and tissues in a model ascidian Ciona savignyi
Reference gene selection for quantitative gene expression studies during biological invasions: A test on multiple genes and tissues in a model ascidian Ciona savignyi
复制标题
生物入侵期间定量基因表达研究的参考基因选择：对模型海鞘 Ciona savignyi 中多个基因和组织的测试
DOI：
10.1016/j.gene.2015.09.066
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发表时间：
2016-01-15
期刊：
影响因子：
3.5
通讯作者：
Zhan, Aibin
中科院分区：
文献类型：
--
作者：
Huang, Xuena;Gao, Yangchun;Zhan, Aibin
由于入侵物种成功地在广泛的截然不同的当地环境中定居，它们为研究物种与快速变化的环境之间的相互作用提供了一个很好的机会。基因表达是快速适应当地环境的主要和关键机制之一。在这里，我们的目标是选择参考基因的定量基因表达分析的基础上定量实时PCR（qRT-PCR）的模式入侵海鞘，海鞘。我们分析了10个候选参考基因的稳定性在三个组织（虹吸管，咽和肠）在两个关键的环境压力（温度和盐度）在海洋领域的基础上的三个程序（geNorm，Normandy和Δ Ct方法）。我们的研究结果表明，三种方法之间的稳定性排名只有微小的差异。使用不同的单个参考基因可能会影响数据解释，而多个参考基因可以最大限度地减少可能的误差。因此，推荐了不同组织的参考基因组合：温度胁迫下，虹吸管的最佳参考基因组合为RPS 15和RPL 17，盐胁迫下的最佳参考基因组合为RPL 17、UBQ和TubA;对于肠道，UBQ、RPS 15和RPL 17是两种处理下最可靠的参考基因。我们的研究结果表明，在不同的环境压力下，不同组织的参考基因的选择和测试的必要性。这里获得的结果有望揭示基因表达介导的入侵成功的机制，使用C savignyi作为模式物种。(C)2015 Elsevier B. V.版权所有。
As invasive species have successfully colonized a wide range of dramatically different local environments, they offer a good opportunity to study interactions between species and rapidly changing environments. Gene expression represents one of the primary and crucial mechanisms for rapid adaptation to local environments. Here, we aim to select reference genes for quantitative gene expression analysis based on quantitative Real-Time PCR (qRT-PCR) for a model invasive ascidian, Ciona savignyi. We analyzed the stability of ten candidate reference genes in three tissues (siphon, pharynx and intestine) under two key environmental stresses (temperature and salinity) in the marine realm based on three programs (geNorm, NormFinder and delta Ct method). Our results demonstrated only minor difference for stability rankings among the three methods. The use of different single reference gene might influence the data interpretation, while multiple reference genes could minimize possible errors. Therefore, reference gene combinations were recommended for different tissues - the optimal reference gene combination for siphon was RPS15 and RPL17 under temperature stress, and RPL17, UBQ and TubA under salinity treatment; for pharynx, TubB, TubA and RPL17 were the most stable genes under temperature stress, while TubB, TubA and UBQ were the best under salinity stress; for intestine, UBQ, RPS15 and RPL17 were the most reliable reference genes under both treatments. Our results suggest that the necessity of selection and test of reference genes for different tissues under varying environmental stresses. The results obtained here are expected to reveal mechanisms of gene expression-mediated invasion success using C savignyi as a model species. (C) 2015 Elsevier B.V. All rights reserved.