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Evaluation of endogenous references for gene expression profiling in different tissues of the oriental fruit fly Bactrocera dorsalis (Diptera: Tephritidae)

机译:评价东方果蝇小实蝇(Bactrocera dorsalis)(双翅目:蝇科)不同组织中基因表达谱的内源性参考

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摘要

BackgroundQuantitative real-time reverse transcriptase PCR (RT-qPCR) has been widely used for quantification of mRNA as a way to determine key genes involved in different biological processes. For accurate gene quantification analysis, normalization of RT-qPCR data is absolutely essential. To date, normalization is most frequently achieved by the use of internal controls, often referred to as reference genes. However, several studies have shown that the reference genes used for the quantification of mRNA expression can be affected by the experimental set-up or cell type resulting in variation of the expression level of these key genes. Therefore, the evaluation of reference genes is critical for gene expression profiling, which is often neglected in gene expression studies of insects. For this purpose, ten candidate reference genes were investigated in three different tissues (midgut, Malpighian tubules, and fat body) of the oriental fruit fly, Bactrocera dorsalis (Hendel).
机译:背景技术实时定量逆转录酶PCR(RT-qPCR)已被广泛用于mRNA定量,以此来确定参与不同生物学过程的关键基因。对于精确的基因定量分析,RT-qPCR数据的标准化绝对必要。迄今为止,标准化通常是通过使用内部对照(通常称为参考基因)来实现的。但是,一些研究表明,用于mRNA表达定量的参考基因会受到实验设置或细胞类型的影响,从而导致这些关键基因的表达水平发生变化。因此,参考基因的评估对于基因表达谱分析至关重要,而在昆虫的基因表达研究中常常忽略了这一点。为此,在东方实蝇Bactrocera dorsalis(Hendel)的三个不同组织(中肠,马尔皮基小管和脂肪体)中研究了十个候选参考基因。

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