Selection of Reference Genes in Transcription Analysis of Gene Expression of the Mandarin Fish, Siniperca chuasti.
At present, transcription analysis of gene expression commonly uses housekeeping genes as control for normalization. In this study, the expression levels of three housekeeping genes including GAPDH, beta-actin, and 18S rRNA in six tissues and five developmental stages of the Mandarin fish Siniperca chuatsi were assayed with quantitative real-time PCR (qPCR). Differences in expression levels were analyzed using geNorm program. The results demonstrate that beta-actin is the most stable gene at developmental stages and GAPDH is the most stable in different tissues. While 18S rRNA expression during development is differentially regulated, which indicates it is suitable as an internal control for gene expression normalization at the developmental level. Overall, the data suggest that the two most stable housekeeping genes are enough to accurately calibrate gene expression in S. chuatsi. The significance of this study provided convincing references and methodology for housekeeping gene selection and normalization in gene expression analysis with regular PCR or qPCR.
Zhou
Rui-Xue
RX
College of Biological Engineering and Environmental Sciences, Changsha University, Changsha 410003, China.
Meng
Tao
T
Meng
Hai-Bo
HB
Cheng
Dun-Xue
DX
Bin
Shi-Yu
SY
Cheng
Jia
J
Fu
Gui-Hong
GH
Chu
Wu-Ying
WY
Zhang
Jian-She
JS
D000199:Actins; D000818:Animals; D015870:Gene Expression; D020869:Gene Expression Profiling; D010473:Perciformes / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000199:Actins