β-Actin is a useful internal control for tissue-specific gene expression studies using quantitative real-time PCR in the half-smooth tongue sole Cynoglossus semilaevis challenged with LPS or Vibrio anguillarum
β-Actin is a useful internal control for tissue-specific gene expression studies using quantitative real-time PCR in the half-smooth tongue sole Cynoglossus semilaevis challenged with LPS or Vibrio anguillarum
复制标题
DOI：
10.1016/j.fsi.2010.02.021
复制
发表时间：
2010-07-01
影响因子：
4.7
通讯作者：
Zhang, Yuanyuan
中科院分区：
文献类型：
--
作者：
Li, Zhaojie;Yang, Lijun;Zhang, Yuanyuan
-actin经常被用作内部控制（基因）或内务管理基因，以使不同样品之间靶基因或mRNA水平的表达正常化。然而，-actin的表达已被证明受到样品类型和实验条件的影响。β -肌动蛋白是否可以作为半光滑舌底的内参基因，目前尚不明确。在本研究中，我们利用实时荧光定量PCR技术，评估了半半C在受到抗原（即脂多糖（LPS）或鳗弧菌）攻击时的组织特异性β -肌动蛋白基因表达模式。根据重组质粒生成的标准曲线进行实时PCR。β -肌动蛋白在处理和未处理的组织样本中的表达没有显著差异，因此我们认为β -肌动蛋白可以作为基于实时PCR的基因表达定量研究的可靠内参基因。(C) 2010 Elsevier Ltd.版权所有。
beta-actin has been frequently used as an internal control (gene) or as a housekeeping gene to normalize the expression of the target gene(s) or mRNA levels between different samples However, the beta-actin expression has been shown to be influenced by the sample type and experimental conditions. beta-actin could be used as a reference gene for the half-smooth tongue sole Cynoglossus semilaevis remains ill-defined. Here we evaluate the tissue-specific beta-actin gene expression pattern in C semilaevis when challenged with antigenic agents namely, lipopolysaccharide (LPS) or Vibrio anguillarum employing absolute quantitative real-time PCR. The real-time PCR was performed based on the standard curve generated from recombinant plasmids. No significant differences in beta-actin expression were found between treated and untreated tissue samples We thus conclude that beta-actin could be used as a reliable internal reference gene for real-time PCR based quantitation of gene expression studies in various tissue samples of C semilaevis challenged with LPS or pathogenic bacteria. (C) 2010 Elsevier Ltd. All rights reserved.