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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
PeerJ Inc.
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4797766/ https://www.ncbi.nlm.nih.gov/pubmed/26998411 http://dx.doi.org/10.7717/peerj.1810 |
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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 |
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 |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-4797766 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | PeerJ Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-47977662016-03-18 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 PeerJ Marine Biology 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. PeerJ Inc. 2016-03-15 /pmc/articles/PMC4797766/ /pubmed/26998411 http://dx.doi.org/10.7717/peerj.1810 Text en ©2016 Chen et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited. |
spellingShingle | Marine Biology 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) |
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 | Marine Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4797766/ https://www.ncbi.nlm.nih.gov/pubmed/26998411 http://dx.doi.org/10.7717/peerj.1810 |
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