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 ( Delphina...
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crpeerj:10.7717/peerj.1810 2024-09-15T17:58:57+00:00 Selection of reference genes for RT-qPCR studies in blood of beluga whales ( Delphinapterus leucas) Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Wu, Yeong-Huey Li, Tsung-Hsien Leu, Ming-Yih Chang, Wen-Been Yang, Wei Cheng 2016 http://dx.doi.org/10.7717/peerj.1810 https://peerj.com/articles/1810.pdf https://peerj.com/articles/1810.xml https://peerj.com/articles/1810.html en eng PeerJ http://creativecommons.org/licenses/by/4.0/ PeerJ volume 4, page e1810 ISSN 2167-8359 journal-article 2016 crpeerj https://doi.org/10.7717/peerj.1810 2024-07-30T04:13:47Z 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. Article in Journal/Newspaper Beluga Beluga* Delphinapterus leucas PeerJ Publishing PeerJ 4 e1810 |
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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. |
format |
Article in Journal/Newspaper |
author |
Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Wu, Yeong-Huey Li, Tsung-Hsien Leu, Ming-Yih Chang, Wen-Been Yang, Wei Cheng |
spellingShingle |
Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Wu, Yeong-Huey Li, Tsung-Hsien Leu, Ming-Yih Chang, Wen-Been Yang, Wei Cheng Selection of reference genes for RT-qPCR studies in blood of beluga whales ( Delphinapterus leucas) |
author_facet |
Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Wu, Yeong-Huey Li, Tsung-Hsien Leu, Ming-Yih Chang, Wen-Been Yang, Wei Cheng |
author_sort |
Chen, I-Hua |
title |
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_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_sort |
selection of reference genes for rt-qpcr studies in blood of beluga whales ( delphinapterus leucas) |
publisher |
PeerJ |
publishDate |
2016 |
url |
http://dx.doi.org/10.7717/peerj.1810 https://peerj.com/articles/1810.pdf https://peerj.com/articles/1810.xml https://peerj.com/articles/1810.html |
genre |
Beluga Beluga* Delphinapterus leucas |
genre_facet |
Beluga Beluga* Delphinapterus leucas |
op_source |
PeerJ volume 4, page e1810 ISSN 2167-8359 |
op_rights |
http://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.7717/peerj.1810 |
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PeerJ |
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4 |
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e1810 |
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1810435905575452672 |