Validation of zebrafish (Danio rerio) reference genes for quantitative real-time RT-PCR normalization
Validation of zebrafish (Danio rerio) reference genes for quantitative real-time RT-PCR normalization
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DOI：
10.1111/j.1745-7270.2007.00283.x
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发表时间：
2007-05-01
影响因子：
3.7
通讯作者：
Love, Donald R.
中科院分区：
文献类型：
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作者：
Tang, Rongying;Dodd, Andrew;Love, Donald R.
定量实时逆转录聚合酶链式反应(qRT-PCR)的标准化对于获得准确的基因表达数据具有重要意义。QRT-PCR归一化最常见的方法是使用参考基因或看家基因。然而，有新的证据表明，即使是参考基因也可以在不同的条件下受到调控。QRT-PCR只是最近才被用于斑马鱼基因表达的研究，还没有一组经过验证的参考基因。这项研究描述了9个可能的参考基因在斑马鱼胚胎发育过程中和斑马鱼组织面板中的表达。所有9个参考基因都表现出可变表达。β-肌动蛋白、EF1α和RPL13α基因构成了斑马鱼发育时程研究的参考基因板，而EF1α、RPL13α和18S rRNA基因更适合作为斑马鱼组织分析的参考基因板。重要的是，斑马鱼GAPDH基因似乎不适合作为这两种类型研究的参考基因。
The normalization of quantitative real time RT-PCR (qRT-PCR) is important to obtain accurate gene expression data. The most common method for qRT-PCR normalization is to use reference, or house keeping genes. However, there is emerging evidence that even reference genes can be regulated under different conditions. qRT-PCR has only recently been used in terms of zebrafish gene expression studies and there is no validated set of reference genes. This study characterizes the expression of nine possible reference genes during zebrafish embryonic development and in a zebrafish tissue panel. All nine reference genes exhibited variable expression. The beta-actin, EF1 alpha and Rpl13 alpha genes comprise a validated reference gene panel for zebrafish developmental time course studies, and the EF1 alpha, Rpl13 alpha and 18S rRNA genes are more suitable as a reference gene panel for zebrafish tissue analysis. Importantly, the zebrafish GAPDH gene appears unsuitable as reference gene for both types of studies.