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Verification of suitable and reliable reference genes for quantitative real-time PCR during adipogenic differentiation in porcine intramuscular stromal-vascular cells

Published online by Cambridge University Press:  19 January 2016

X. Li ,
K. Huang ,
F. Chen ,
W. Li ,
S. Sun ,
X.-E. Shi  and
G. Yang
X. Li
Affiliation:
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, P. R.China
K. Huang
Affiliation:
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, P. R.China
F. Chen
Affiliation:
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, P. R.China
W. Li
Affiliation:
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, P. R.China
S. Sun
Affiliation:
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, P. R.China
X.-E. Shi
Affiliation:
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, P. R.China
G. Yang*
Affiliation:
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, P. R.China
*
†E-mail: gsyang999@hotmail.com
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Abstract

Intramuscular fat (IMF) is an important trait influencing meat quality, and intramuscular stromal-vascular cell (MSVC) differentiation is a key factor affecting IMF deposition. Quantitative real-time PCR (qPCR) is often used to screen the differentially expressed genes during differentiation of MSVCs, where proper reference genes are essential. In this study, we assessed 31 of previously reported reference genes for their expression suitability in porcine MSVCs derived form longissimus dorsi with qPCR. The expression stability of these genes was evaluated using NormFinder, geNorm and BestKeeper algorithms. NormFinder and geNorm uncovered ACTB, ALDOA and RPS18 as the most three stable genes. BestKeeper identified RPL13A, SSU72 and DAK as the most three stable genes. GAPDH was found to be the least stable gene by all of the three software packages, indicating it is not an appropriate reference gene in qPCR assay. These results might be helpful for further studies in pigs that explore the molecular mechanism underlying IMF deposition.

Keywords

reference genequantitative real-time PCRpigintramuscular stromal-vascular cellsintramuscular fat
Type
Research Article
Information
animal , Volume 10 , Issue 6 , June 2016 , pp. 947 - 952
Copyright
© The Animal Consortium 2016 

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Footnotes

a

Both authors contributed equally to this work.

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