beta-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.
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. If 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.
Li
Zhaojie
Z
The Technology Center of Weihai Entry-exit Inspection and Quarantine Bureau, Weihai 264205, PR China.
Yang
Lijun
L
Wang
Jing
J
Shi
Wenchun
W
Pawar
Ravindra Ajit
RA
Liu
Yumin
Y
Xu
Chenggang
C
Cong
Weihong
W
Hu
Qiaoru
Q
Lu
Taiyi
T
Xia
Fan
F
Guo
Wei
W
Zhao
Meng
M
Zhang
Yuanyuan
Y
D000199:Actins / Q000235:genetics / Q000276:immunology*; D000818:Animals; D005412:Flatfishes / Q000235:genetics / Q000276:immunology*; D018628:Gene Dosage; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards; D008070:Lipopolysaccharides / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D014733:Vibrio / Q000276:immunology*
D000199:Actins; D008070:Lipopolysaccharides; D012313:RNA