Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety
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
Affiliations
Kelly J. Lozano-Villegas
Immunobiology and Pathogenesis Research Group, Faculty of Veterinary Medicine and Zootechnics, University of Tolima, Altos the Santa Helena, A.A 546, Ibagué 730006299, Tolima, Colombia
Roy Rodríguez-Hernández
Poultry Research Group, Laboratory of Immunology and Molecular Biology, Faculty of Veterinary Medicine and Zootechnics, University of Tolima, Altos the Santa Helena, A.A 546, Ibagué 730006299, Tolima, Colombia
María P. Herrera-Sánchez
Immunobiology and Pathogenesis Research Group, Faculty of Veterinary Medicine and Zootechnics, University of Tolima, Altos the Santa Helena, A.A 546, Ibagué 730006299, Tolima, Colombia
Heinner F. Uribe-García
Immunobiology and Pathogenesis Research Group, Faculty of Veterinary Medicine and Zootechnics, University of Tolima, Altos the Santa Helena, A.A 546, Ibagué 730006299, Tolima, Colombia
Juan S. Naranjo-Gómez
Immunobiology and Pathogenesis Research Group, Faculty of Veterinary Medicine and Zootechnics, University of Tolima, Altos the Santa Helena, A.A 546, Ibagué 730006299, Tolima, Colombia
Rafael J. Otero-Arroyo
Grupo de Investigación en Reproducción y Mejoramiento Genético Animal, Facultad de Ciencias Agropecuarias, Universidad de Sucre, Sincelejo 700001, Sucre, Colombia
Iang S. Rondón-Barragán
Immunobiology and Pathogenesis Research Group, Faculty of Veterinary Medicine and Zootechnics, University of Tolima, Altos the Santa Helena, A.A 546, Ibagué 730006299, Tolima, Colombia
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 (actb, ywhaz, b2m, sdha, and 18s rRNA) at different physiological stages (alert and emergency) in three different cattle breeds. In this study, five genes (actb, ywhaz, b2m, sdha, and 18s rRNA) 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 actb, 18SrRNA, and b2m 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, actb, b2m, and ywhaz were the most stable reference genes for the Romosinuano breed.
