title: Selection of appropriate reference genes for RT-qPCR analysis in Berkshire, Duroc, Landrace, and Yorkshire pigs.
abstract: Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is the most reliable molecular biology technique for assessment of mRNA expression levels. However, to obtain the accurate RT-qPCR results, the expression levels of genes of interest should be normalized with appropriate reference genes and optimal numbers of reference genes. In this study, we assessed the expression stability of 15 well-known candidate reference genes (ACTB, ALDOA, B2M, GAPDH, HPAR1, HSPCB, PGK1, POLR2G, PPIA, RPL4, RPS18, SDHA, TBP, TOP2B, and YWHAZ) in seven body tissues (liver, lung, kidney, spleen, stomach, small intestine, and large intestine) of Berkshire, Landrace, Duroc, and Yorkshire pigs using three excel-based programs, geNorm, NormFinder, and BestKeeper. Combination analysis of these three programs showed that the stable and appropriate reference genes are PPIA, TBP, and HSPCB in Berkshire pigs; PPIA, TBP, RPL4, and RPS18 in Landrace pigs; PPIA and TBP in Duroc pigs; and PPIA, TOP2B, RPL4, and RPS18 in Yorkshire pigs. Because the four pig breeds had different suitable reference genes, the selection of appropriate reference genes is essential in RT-qPCR analyses. Taken together, our data could help to select reliable reference genes for the normalization of expression levels of various target genes in pigs.
authors.affiliation: National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungbuk 363-883, Republic of Korea. Electronic address: sjpark@gntech.ac.kr.
authors.forename: Sang-Je
authors.initials: SJ
authors.lastname: Park
authors.affiliation: Swine Science and Technology Center, Gyeongnam National University of Science and Technology, Jinju 660-758, Republic of Korea.
authors.forename: Seul Gi
authors.initials: SG
authors.lastname: Kwon
authors.affiliation: Swine Science and Technology Center, Gyeongnam National University of Science and Technology, Jinju 660-758, Republic of Korea.
authors.forename: Jung Hye
authors.initials: JH
authors.lastname: Hwang
authors.affiliation: Swine Science and Technology Center, Gyeongnam National University of Science and Technology, Jinju 660-758, Republic of Korea.
authors.forename: Da Hye
authors.initials: DH
authors.lastname: Park
authors.affiliation: Swine Science and Technology Center, Gyeongnam National University of Science and Technology, Jinju 660-758, Republic of Korea.
authors.forename: Tae Wan
authors.initials: TW
authors.lastname: Kim
authors.affiliation: Swine Science and Technology Center, Gyeongnam National University of Science and Technology, Jinju 660-758, Republic of Korea. Electronic address: cwkim@gntech.ac.kr.
authors.forename: Chul Wook
authors.initials: CW
authors.lastname: Kim
mesh_terms: D000818:Animals; D001483:Base Sequence; D008969:Molecular Sequence Data; D060888:Real-Time Polymerase Chain Reaction; D012015:Reference Standards; D034421:Sus scrofa / Q000145:classification* / Q000235:genetics*; D059467:Transcriptome
keywords: Berkshire; Duroc; Landrace; Normalization; RT-qPCR; Reference gene; Yorkshire