Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas)
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 (Delphinap...
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| Format: | Article |
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PeerJ Inc.
2016-03-01
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| Online Access: | https://peerj.com/articles/1810.pdf |
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| author | I-Hua Chen Jiann-Hsiung Wang Shih-Jen Chou Yeong-Huey Wu Tsung-Hsien Li Ming-Yih Leu Wen-Been Chang Wei Cheng Yang |
| author_facet | I-Hua Chen Jiann-Hsiung Wang Shih-Jen Chou Yeong-Huey Wu Tsung-Hsien Li Ming-Yih Leu Wen-Been Chang Wei Cheng Yang |
| author_sort | I-Hua Chen |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T06:54:09Z |
| format | Article |
| id | doaj.art-6a2da766d57047ca85a01e4262ab00b2 |
| institution | Directory Open Access Journal |
| issn | 2167-8359 |
| language | English |
| last_indexed | 2024-03-09T06:54:09Z |
| publishDate | 2016-03-01 |
| publisher | PeerJ Inc. |
| record_format | Article |
| series | PeerJ |
| spelling | doaj.art-6a2da766d57047ca85a01e4262ab00b22023-12-03T10:07:36ZengPeerJ Inc.PeerJ2167-83592016-03-014e181010.7717/peerj.1810Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas)I-Hua Chen0Jiann-Hsiung Wang1Shih-Jen Chou2Yeong-Huey Wu3Tsung-Hsien Li4Ming-Yih Leu5Wen-Been Chang6Wei Cheng Yang7Department of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan, ROCDepartment of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan, ROCDepartment of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan, ROCDepartment of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan, ROCDepartment of Biology, National Museum of Marine Biology and Aquarium, Pingtung, Taiwan, ROCDepartment of Biology, National Museum of Marine Biology and Aquarium, Pingtung, Taiwan, ROCDepartment of Biology, National Museum of Marine Biology and Aquarium, Pingtung, Taiwan, ROCDepartment of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan, ROCReverse 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.https://peerj.com/articles/1810.pdfBelugaHousekeepingQuantitative PCRGene expression |
| spellingShingle | I-Hua Chen Jiann-Hsiung Wang Shih-Jen Chou Yeong-Huey Wu Tsung-Hsien Li Ming-Yih Leu Wen-Been Chang Wei Cheng Yang Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas) PeerJ Beluga Housekeeping Quantitative PCR Gene expression |
| title | Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas) |
| title_full | Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas) |
| title_fullStr | Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas) |
| title_full_unstemmed | Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas) |
| title_short | Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas) |
| title_sort | selection of reference genes for rt qpcr studies in blood of beluga whales delphinapterus leucas |
| topic | Beluga Housekeeping Quantitative PCR Gene expression |
| url | https://peerj.com/articles/1810.pdf |
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