Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety

Real-time PCR is widely used to study the relative abundance of mRNA due to its specificity, sensitivity, and repeatability quantification. However, relative quantification requires a reference gene, which should be stable in its expression, showing lower variation by experimental conditions or tiss...

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Autores principales:	Kelly J. Lozano-Villegas, Roy Rodríguez-Hernández, María P. Herrera-Sánchez, Heinner F. Uribe-García, Juan S. Naranjo-Gómez, Rafael J. Otero-Arroyo, Iang S. Rondón-Barragán
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	cattle farm animals environmental conditions heat stress mRNA genes normalization Veterinary medicine SF600-1100 Zoology QL1-991
Acceso en línea:	https://doaj.org/article/992f338727d54b0692341233924a89c5
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spelling	oai:doaj.org-article:992f338727d54b0692341233924a89c52021-11-25T16:14:34ZIdentification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety10.3390/ani111130732076-2615https://doaj.org/article/992f338727d54b0692341233924a89c52021-10-01T00:00:00Zhttps://www.mdpi.com/2076-2615/11/11/3073https://doaj.org/toc/2076-2615Real-time PCR is widely used to study the relative abundance of mRNA due to its specificity, sensitivity, and repeatability quantification. However, relative quantification requires a reference gene, which should be stable in its expression, showing lower variation by experimental conditions or tissues. The aim of this study was to evaluate the stability of the expression of five commonly used reference genes <i>(actb</i>, <i>ywhaz</i>, <i>b2m</i>, <i>sdha</i>, and <i>18s rRNA</i>) at different physiological stages (alert and emergency) in three different cattle breeds. In this study, five genes (<i>actb</i>, <i>ywhaz</i>, <i>b2m</i>, <i>sdha</i>, and <i>18s rRNA</i>) were selected as candidate reference genes for expression studies in the whole blood from three cattle breeds (Romosinuano, Gyr, and Brahman) under heat stress conditions. The transcription stability of the candidate reference genes was evaluated using geNorm and NormFinder. The results showed that <i>actb</i>, <i>18SrRNA</i>, and <i>b2m</i> expression were the most stable reference genes for whole blood of Gyr and Brahman breeds under two states of livestock weather safety (alert and emergency). Meanwhile, <i>actb</i>, <i>b2m</i>, and <i>ywhaz</i> were the most stable reference genes for the Romosinuano breed.Kelly J. Lozano-VillegasRoy Rodríguez-HernándezMaría P. Herrera-SánchezHeinner F. Uribe-GarcíaJuan S. Naranjo-GómezRafael J. Otero-ArroyoIang S. Rondón-BarragánMDPI AGarticlecattlefarm animalsenvironmental conditionsheat stressmRNAgenes normalizationVeterinary medicineSF600-1100ZoologyQL1-991ENAnimals, Vol 11, Iss 3073, p 3073 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	cattle farm animals environmental conditions heat stress mRNA genes normalization Veterinary medicine SF600-1100 Zoology QL1-991
spellingShingle	cattle farm animals environmental conditions heat stress mRNA genes normalization Veterinary medicine SF600-1100 Zoology QL1-991 Kelly J. Lozano-Villegas Roy Rodríguez-Hernández María P. Herrera-Sánchez Heinner F. Uribe-García Juan S. Naranjo-Gómez Rafael J. Otero-Arroyo Iang S. Rondón-Barragán Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety
description	Real-time PCR is widely used to study the relative abundance of mRNA due to its specificity, sensitivity, and repeatability quantification. However, relative quantification requires a reference gene, which should be stable in its expression, showing lower variation by experimental conditions or tissues. The aim of this study was to evaluate the stability of the expression of five commonly used reference genes <i>(actb</i>, <i>ywhaz</i>, <i>b2m</i>, <i>sdha</i>, and <i>18s rRNA</i>) at different physiological stages (alert and emergency) in three different cattle breeds. In this study, five genes (<i>actb</i>, <i>ywhaz</i>, <i>b2m</i>, <i>sdha</i>, and <i>18s rRNA</i>) were selected as candidate reference genes for expression studies in the whole blood from three cattle breeds (Romosinuano, Gyr, and Brahman) under heat stress conditions. The transcription stability of the candidate reference genes was evaluated using geNorm and NormFinder. The results showed that <i>actb</i>, <i>18SrRNA</i>, and <i>b2m</i> expression were the most stable reference genes for whole blood of Gyr and Brahman breeds under two states of livestock weather safety (alert and emergency). Meanwhile, <i>actb</i>, <i>b2m</i>, and <i>ywhaz</i> were the most stable reference genes for the Romosinuano breed.
format	article
author	Kelly J. Lozano-Villegas Roy Rodríguez-Hernández María P. Herrera-Sánchez Heinner F. Uribe-García Juan S. Naranjo-Gómez Rafael J. Otero-Arroyo Iang S. Rondón-Barragán
author_facet	Kelly J. Lozano-Villegas Roy Rodríguez-Hernández María P. Herrera-Sánchez Heinner F. Uribe-García Juan S. Naranjo-Gómez Rafael J. Otero-Arroyo Iang S. Rondón-Barragán
author_sort	Kelly J. Lozano-Villegas
title	Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety
title_short	Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety
title_full	Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety
title_fullStr	Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety
title_full_unstemmed	Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety
title_sort	identification of reference genes for expression studies in the whole-blood from three cattle breeds under two states of livestock weather safety
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/992f338727d54b0692341233924a89c5
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Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety

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