Evaluation of housekeeping genes for normalizing real-time quantitative PCR assays in pig skeletal muscle at multiple developmental stages

被引:29
作者
Wang, Yueying [1 ]
Zhao, Yuqiang [1 ]
Li, Ji [1 ]
Liu, Huijing [1 ]
Ernst, Catherine W. [2 ]
Liu, Xiaoran [3 ]
Liu, Guorong [1 ]
Xi, Yu [1 ]
Lei, Minggang [1 ]
机构
[1] Huazhong Cent China Agr Univ, Coll Anim Sci & Technol, Key Lab Agr Anim Genet Breeding & Reprod, Minist Educ,Minist Agr,Key Lab Swine Genet & Bree, Wuhan 430070, Hubei, Peoples R China
[2] Michigan State Univ, Dept Anim Sci, E Lansing, MI 48824 USA
[3] Huazhong Agr Univ, Coll Anim Sci & Technol, China Educ Minist, Key Lab Agr Anim Genet Breeding & Reprod, Wuhan 430070, Peoples R China
关键词
Reference gene; q-PCR; Skeletal muscle; Multiple developmental stages; Pig; POLYMERASE-CHAIN-REACTION; MESSENGER-RNA LEVELS; RT-PCR; BETA-ACTIN; PREIMPLANTATION EMBRYOS; EXPRESSION; QUANTIFICATION; VALIDATION; SELECTION; TISSUE;
D O I
10.1016/j.gene.2015.04.016
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Quantitative PCR (qPCR) requires a constantly expressed housekeeping gene as an internal control, the expression of which is similar in different biological samples. In the present study, we evaluated the applicability and compared the consistency of the gene expression of 16 reference genes, i.e., 10 common and 6 candidate genes, through qPCR assays in pig skeletal muscles at multiple developmental stages. The stability of these 16 potential reference genes was investigated using the geNorm and NormFinder methods. Our results indicated that DRAP1 and RNF7 were the most appropriate combination to normalize gene expression in the Yorkshire samples, the combination of DRAP1 and WSB2 were appropriate in the Tongcheng samples, H3F3A and DRAM in prenatal periods, DRAP1 and RNF7 in postnatal periods, and the combination of DRAP1 and WSB2 was most suitable for accurate normalization in whole samples. This result provides a strong basis for similar studies in pigs that explore the molecular mechanisms underlying skeletal muscle development. (C) 2015 Elsevier B.V. All rights reserved.
引用
收藏
页码:235 / 241
页数:7
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