title: Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas).
abstract: Reverse transcription quantitative PCR (RT-qPCR) is used for research in gene expression, and it is vital to choose appropriate housekeeping genes (HKGs) as reference genes to obtain correct results. The purpose of this study is to determine stably expressed HKGs in blood of beluga whales (Delphinapterus leucas) that can be the appropriate reference genes in relative quantification in gene expression research. Sixty blood samples were taken from four beluga whales. Thirteen candidate HKGs (ACTB, B2M, GAPDH, HPRT1, LDHB, PGK1, RPL4, RPL8, RPL18, RPS9, RPS18, TFRC, YWHAZ) were tested using RT-qPCR. The stability values of the HKGs were determined by four different algorithms. Comprehensive analysis of the results revealed that RPL4, PGK1 and ACTB are strongly recommended for use in future RT-qPCR studies in beluga blood samples. This research provides recommendation of reference gene selection, which may contribute to further mRNA relative quantification research in the peripheral blood leukocytes in captive cetaceans. The gene expression assessment of the immune components in blood have the potential to serve as an important approach to evaluating cetacean health influenced by environmental insults.
authors.affiliation: Department of Veterinary Medicine, National Chiayi University , Chiayi , Taiwan, ROC.
authors.forename: I-Hua
authors.initials: IH
authors.lastname: Chen
authors.affiliation: Department of Veterinary Medicine, National Chiayi University , Chiayi , Taiwan, ROC.
authors.forename: Jiann-Hsiung
authors.initials: JH
authors.lastname: Wang
authors.affiliation: Department of Veterinary Medicine, National Chiayi University , Chiayi , Taiwan, ROC.
authors.forename: Shih-Jen
authors.initials: SJ
authors.lastname: Chou
authors.affiliation: Department of Veterinary Medicine, National Pingtung University of Science and Technology , Pingtung , Taiwan, ROC.
authors.forename: Yeong-Huey
authors.initials: YH
authors.lastname: Wu
authors.affiliation: Department of Biology, National Museum of Marine Biology and Aquarium , Pingtung , Taiwan, ROC.
authors.forename: Tsung-Hsien
authors.initials: TH
authors.lastname: Li
authors.affiliation: Department of Biology, National Museum of Marine Biology and Aquarium, Pingtung, Taiwan, ROC; Graduate Institute of Marine Biology, National Dong Hwa University, Pingtung, Taiwan, ROC.
authors.forename: Ming-Yih
authors.initials: MY
authors.lastname: Leu
authors.affiliation: Department of Biology, National Museum of Marine Biology and Aquarium , Pingtung , Taiwan, ROC.
authors.forename: Wen-Been
authors.initials: WB
authors.lastname: Chang
authors.affiliation: Department of Veterinary Medicine, National Chiayi University , Chiayi , Taiwan, ROC.
authors.forename: Wei Cheng
authors.initials: WC
authors.lastname: Yang
keywords: Beluga; Gene expression; Housekeeping; Quantitative PCR