4.7 Article

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

Journal

FISH & SHELLFISH IMMUNOLOGY
Volume 29, Issue 1, Pages 89-93

Publisher

ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.fsi.2010.02.021

Keywords

beta-Actin; Absolute quantitation; Standard curve; Cynoglossus semilaevis; Lipopolysaccharide; Vibrio anguillarum

Funding

  1. Shandong Province Natural Science Foundation [ZR2009EQ009]

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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.

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