Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Revista Brasileira de Zootecnia (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-35982022000100402 |
Resumo: | ABSTRACT The objective of the present study was to evaluate the stability of candidate reference genes and select the genes that can be used for normalizing real-time polymerase chain reaction (PCR) in the liver, skeletal muscle, and jejunum tissues of Nellore or Nellore × Angus steers fed different diets. Fourteen purebred and 14 crossbred steers were used, in which half of the animals of each genetic group received a diet containing whole shelled corn (WSC) and the other half whole shelled corn and sugarcane bagasse (WSCB). Stability was analyzed by the RefFinder program. To validate the selection of candidate reference genes, the expression of target genes was evaluated using the different groups of reference genes. The most stable genes were 18S, ACTB, and CASC3 for skeletal muscle; HMBS, ACTB, and 18S for the liver; and GAPDH, ACTB, and CASC3 for the jejunum, regardless of breed and diet provided. Possible errors caused in data analyses were clarified comparing the more and less stable genes as reference for normalization of the target genes FASN, ACOX, SCD1, MGAM, and SLC2A1. The use of the more stable and less stable sets of reference genes may lead to different conclusions in respect to the expression profile of the target studied gene. The selection of more suitable reference genes for each experiment is of utmost importance to ensure the reliability of gene expression studies so that they can be applied in practice. |
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Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattlebovinehousekeeping genesnormalizationRT-qPCRtissuesABSTRACT The objective of the present study was to evaluate the stability of candidate reference genes and select the genes that can be used for normalizing real-time polymerase chain reaction (PCR) in the liver, skeletal muscle, and jejunum tissues of Nellore or Nellore × Angus steers fed different diets. Fourteen purebred and 14 crossbred steers were used, in which half of the animals of each genetic group received a diet containing whole shelled corn (WSC) and the other half whole shelled corn and sugarcane bagasse (WSCB). Stability was analyzed by the RefFinder program. To validate the selection of candidate reference genes, the expression of target genes was evaluated using the different groups of reference genes. The most stable genes were 18S, ACTB, and CASC3 for skeletal muscle; HMBS, ACTB, and 18S for the liver; and GAPDH, ACTB, and CASC3 for the jejunum, regardless of breed and diet provided. Possible errors caused in data analyses were clarified comparing the more and less stable genes as reference for normalization of the target genes FASN, ACOX, SCD1, MGAM, and SLC2A1. The use of the more stable and less stable sets of reference genes may lead to different conclusions in respect to the expression profile of the target studied gene. The selection of more suitable reference genes for each experiment is of utmost importance to ensure the reliability of gene expression studies so that they can be applied in practice.Sociedade Brasileira de Zootecnia2022-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-35982022000100402Revista Brasileira de Zootecnia v.51 2022reponame:Revista Brasileira de Zootecnia (Online)instname:Sociedade Brasileira de Zootecnia (SBZ)instacron:SBZ10.37496/rbz5120210120info:eu-repo/semantics/openAccessCoelho,Tamara CristinaChalfun-Junior,AntonioBarreto,Horllys GomesDuarte,Marcio de SouzaGarcia,Bruno de OliveiraTeixeira,Priscilla DutraGionbelli,Tathyane Ramalho SantosLadeira,Márcio Machadoeng2022-07-15T00:00:00Zoai:scielo:S1516-35982022000100402Revistahttps://www.rbz.org.br/pt-br/https://old.scielo.br/oai/scielo-oai.php||bz@sbz.org.br|| secretariarbz@sbz.org.br1806-92901516-3598opendoar:2022-07-15T00:00Revista Brasileira de Zootecnia (Online) - Sociedade Brasileira de Zootecnia (SBZ)false |
dc.title.none.fl_str_mv |
Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle |
title |
Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle |
spellingShingle |
Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle Coelho,Tamara Cristina bovine housekeeping genes normalization RT-qPCR tissues |
title_short |
Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle |
title_full |
Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle |
title_fullStr |
Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle |
title_full_unstemmed |
Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle |
title_sort |
Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle |
author |
Coelho,Tamara Cristina |
author_facet |
Coelho,Tamara Cristina Chalfun-Junior,Antonio Barreto,Horllys Gomes Duarte,Marcio de Souza Garcia,Bruno de Oliveira Teixeira,Priscilla Dutra Gionbelli,Tathyane Ramalho Santos Ladeira,Márcio Machado |
author_role |
author |
author2 |
Chalfun-Junior,Antonio Barreto,Horllys Gomes Duarte,Marcio de Souza Garcia,Bruno de Oliveira Teixeira,Priscilla Dutra Gionbelli,Tathyane Ramalho Santos Ladeira,Márcio Machado |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Coelho,Tamara Cristina Chalfun-Junior,Antonio Barreto,Horllys Gomes Duarte,Marcio de Souza Garcia,Bruno de Oliveira Teixeira,Priscilla Dutra Gionbelli,Tathyane Ramalho Santos Ladeira,Márcio Machado |
dc.subject.por.fl_str_mv |
bovine housekeeping genes normalization RT-qPCR tissues |
topic |
bovine housekeeping genes normalization RT-qPCR tissues |
description |
ABSTRACT The objective of the present study was to evaluate the stability of candidate reference genes and select the genes that can be used for normalizing real-time polymerase chain reaction (PCR) in the liver, skeletal muscle, and jejunum tissues of Nellore or Nellore × Angus steers fed different diets. Fourteen purebred and 14 crossbred steers were used, in which half of the animals of each genetic group received a diet containing whole shelled corn (WSC) and the other half whole shelled corn and sugarcane bagasse (WSCB). Stability was analyzed by the RefFinder program. To validate the selection of candidate reference genes, the expression of target genes was evaluated using the different groups of reference genes. The most stable genes were 18S, ACTB, and CASC3 for skeletal muscle; HMBS, ACTB, and 18S for the liver; and GAPDH, ACTB, and CASC3 for the jejunum, regardless of breed and diet provided. Possible errors caused in data analyses were clarified comparing the more and less stable genes as reference for normalization of the target genes FASN, ACOX, SCD1, MGAM, and SLC2A1. The use of the more stable and less stable sets of reference genes may lead to different conclusions in respect to the expression profile of the target studied gene. The selection of more suitable reference genes for each experiment is of utmost importance to ensure the reliability of gene expression studies so that they can be applied in practice. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-01-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-35982022000100402 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-35982022000100402 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.37496/rbz5120210120 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Zootecnia |
publisher.none.fl_str_mv |
Sociedade Brasileira de Zootecnia |
dc.source.none.fl_str_mv |
Revista Brasileira de Zootecnia v.51 2022 reponame:Revista Brasileira de Zootecnia (Online) instname:Sociedade Brasileira de Zootecnia (SBZ) instacron:SBZ |
instname_str |
Sociedade Brasileira de Zootecnia (SBZ) |
instacron_str |
SBZ |
institution |
SBZ |
reponame_str |
Revista Brasileira de Zootecnia (Online) |
collection |
Revista Brasileira de Zootecnia (Online) |
repository.name.fl_str_mv |
Revista Brasileira de Zootecnia (Online) - Sociedade Brasileira de Zootecnia (SBZ) |
repository.mail.fl_str_mv |
||bz@sbz.org.br|| secretariarbz@sbz.org.br |
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1750318154342465536 |