Screening of Reference Genes for qRT-PCR in Bama Minipig (Sus scrofa) Tissues
CHEN Chu-Jie1, PEI Yang-Li1,*, LIU Lu-Lu2, YANG Ya-Lan1, ZHANG Lei-Xia1, LI Hua1, LI Kui1,2
1 Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding/School of Life Sciences and Engineering, Foshan University, Foshan 528225, China; 2 Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Abstract：qRT-PCR is one of the important methods for investigating mRNA expression levels in the cells and tissues. However, the selection of suitable reference genes is crucial to accurately evaluate and normalize the relative expression level of target genes for gene function analysis. This study examined the expression of 9 internal genes (topoisomerase (DNA)Ⅱβ (Top2b), β-actin (Actb), ribosomal protein S18 (Rps18), peptidylprolyl isomerase A (Ppia), 18S ribosomal RNA (18S), TATA-box binding protein (Tbp), hydroxymethylbilane synthase (Hmbs), ribosomal protein L4 (Rpl4) and β-2-microglobulin (B2m) for heart, liver, spleen, lung, kidney, muscle, subcutaneous fat from three 15 d Bama minipigs (Sus scrofa) using qRT-PCR technique, and 3 well-known software packages: geNorm, NormFinder and BestKeeper were used to analyze the results. Rps18, Rpl4, Ppia and Tbp were found to own the highest stability across tissues. Only 2 internal reference genes could standardize the expression of target genes in the same tissue. And in the different tissue, the optimal gene combination was different. The results of this study provide a reference information on Bama minipig to select the most suitable internal reference gene, and it could provide a theoretical basis for the research of gene function about meat quality, litter size, development, reproduction, disease and so on of Bama minipig.
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