| Validation of Zebrafish （Danio redo） Reference Genes for Quantitative Real-time RT-PCR Normalization |
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| 作者姓名： | Tang R Dodd A Lai D McNabb WC Love DR |
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| 作者单位： | [1]School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland 1142, New Zealand [2]Food and Health Group, AgResearch Grasslands, Private Bag 11008, Tennant Drive, Palmerston North, New Zealand |
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| 基金项目： | This work was supported by the grants from the University of Auckland Research Grants Committee, the Maurice and Phyllis Paykel Trust and the Lottery Grants Board of New Zealand |
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| 摘 要： | 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 housekeeping 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 fl-actin, EFlot and Rpll3ot genes comprise a validated reference gene panel for zebrafish developmental time course studies, and the EF1 or, Rpll3α 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.
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| 关 键 词： | 斑马鱼 定量实时RT-PCR 归化法 参照基因 |
| 修稿时间： | 2006-12-132007-03-05 |
Validation of zebrafish (Danio rerio) reference genes for quantitative real-time RT-PCR normalization |
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| Tang R,Dodd A,Lai D,McNabb WC,Love DR.Validation of zebrafish (Danio rerio) reference genes for quantitative real-time RT-PCR normalization[J].Acta Biochimica et Biophysica Sinica,2007,39(5):384-390. |
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| Authors: | Tang Rongying Dodd Andrew Lai Daniel McNabb Warren C Love Donald R |
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| Institution: | School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland 1142, New Zealand. |
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| Abstract: | 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 housekeeping 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, EF1alpha and Rpl13alpha genes comprise a validated reference gene panel for zebrafish developmental time course studies, and the EF1alpha, Rpl13alpha 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. |
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| Keywords: | zebrafish quantitative real-time RT-PCR housekeeping genes GAPDH gene GeNorm |
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