Screening of Reference Genes for RT-qPCR in Chicken Adipose Tissue and Adipocytes

Reverse transcription quantitative real-time PCR is the most commonly used method to detect gene expression levels. In experiments, it is often necessary to correct and standardize the expression level of target genes with reference genes. Therefore, it is very important to select stable reference g...

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Autores principales: Na, Wei, Wang, Yuxiang, Gong, Pengfei, Zhang, Xinyang, Zhang, Ke, Zhang, Hui, Wang, Ning, Li, Hui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8160385/
https://www.ncbi.nlm.nih.gov/pubmed/34054585
http://dx.doi.org/10.3389/fphys.2021.676864
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author Na, Wei
Wang, Yuxiang
Gong, Pengfei
Zhang, Xinyang
Zhang, Ke
Zhang, Hui
Wang, Ning
Li, Hui
author_facet Na, Wei
Wang, Yuxiang
Gong, Pengfei
Zhang, Xinyang
Zhang, Ke
Zhang, Hui
Wang, Ning
Li, Hui
author_sort Na, Wei
collection PubMed
description Reverse transcription quantitative real-time PCR is the most commonly used method to detect gene expression levels. In experiments, it is often necessary to correct and standardize the expression level of target genes with reference genes. Therefore, it is very important to select stable reference genes to obtain accurate quantitative results. Although application examples of reference genes in mammals have been reported, no studies have investigated the use of reference genes in studying the growth and development of adipose tissue and the proliferation and differentiation of preadipocytes in chickens. In this study, GeNorm, a reference gene stability statistical algorithm, was used to analyze the expression stability of 14 candidate reference genes in the abdominal adipose tissue of broilers at 1, 4, and 7 weeks of age, the proliferation and differentiation of primary preadipocytes, as well as directly isolated preadipocytes and mature adipocytes. The results showed that the expression of the TATA box binding protein (TBP) and hydroxymethylbilane synthase (HMBS) genes was most stable during the growth and development of abdominal adipose tissue of broilers, the expression of the peptidylprolyl isomerase A (PPIA) and HMBS genes was most stable during the proliferation of primary preadipocytes, the expression of the TBP and RPL13 genes was most stable during the differentiation of primary preadipocytes, and the expression of the TBP and HMBS genes was most stable in directly isolated preadipocytes and mature adipocytes. These results provide reference bases for accurately detecting the mRNA expression of functional genes in adipose tissue and adipocytes of chickens.
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spelling pubmed-81603852021-05-29 Screening of Reference Genes for RT-qPCR in Chicken Adipose Tissue and Adipocytes Na, Wei Wang, Yuxiang Gong, Pengfei Zhang, Xinyang Zhang, Ke Zhang, Hui Wang, Ning Li, Hui Front Physiol Physiology Reverse transcription quantitative real-time PCR is the most commonly used method to detect gene expression levels. In experiments, it is often necessary to correct and standardize the expression level of target genes with reference genes. Therefore, it is very important to select stable reference genes to obtain accurate quantitative results. Although application examples of reference genes in mammals have been reported, no studies have investigated the use of reference genes in studying the growth and development of adipose tissue and the proliferation and differentiation of preadipocytes in chickens. In this study, GeNorm, a reference gene stability statistical algorithm, was used to analyze the expression stability of 14 candidate reference genes in the abdominal adipose tissue of broilers at 1, 4, and 7 weeks of age, the proliferation and differentiation of primary preadipocytes, as well as directly isolated preadipocytes and mature adipocytes. The results showed that the expression of the TATA box binding protein (TBP) and hydroxymethylbilane synthase (HMBS) genes was most stable during the growth and development of abdominal adipose tissue of broilers, the expression of the peptidylprolyl isomerase A (PPIA) and HMBS genes was most stable during the proliferation of primary preadipocytes, the expression of the TBP and RPL13 genes was most stable during the differentiation of primary preadipocytes, and the expression of the TBP and HMBS genes was most stable in directly isolated preadipocytes and mature adipocytes. These results provide reference bases for accurately detecting the mRNA expression of functional genes in adipose tissue and adipocytes of chickens. Frontiers Media S.A. 2021-05-14 /pmc/articles/PMC8160385/ /pubmed/34054585 http://dx.doi.org/10.3389/fphys.2021.676864 Text en Copyright © 2021 Na, Wang, Gong, Zhang, Zhang, Zhang, Wang and Li. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Physiology
Na, Wei
Wang, Yuxiang
Gong, Pengfei
Zhang, Xinyang
Zhang, Ke
Zhang, Hui
Wang, Ning
Li, Hui
Screening of Reference Genes for RT-qPCR in Chicken Adipose Tissue and Adipocytes
title Screening of Reference Genes for RT-qPCR in Chicken Adipose Tissue and Adipocytes
title_full Screening of Reference Genes for RT-qPCR in Chicken Adipose Tissue and Adipocytes
title_fullStr Screening of Reference Genes for RT-qPCR in Chicken Adipose Tissue and Adipocytes
title_full_unstemmed Screening of Reference Genes for RT-qPCR in Chicken Adipose Tissue and Adipocytes
title_short Screening of Reference Genes for RT-qPCR in Chicken Adipose Tissue and Adipocytes
title_sort screening of reference genes for rt-qpcr in chicken adipose tissue and adipocytes
topic Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8160385/
https://www.ncbi.nlm.nih.gov/pubmed/34054585
http://dx.doi.org/10.3389/fphys.2021.676864
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