Validation of suitable reference genes for quantitative expression analysis by qPCR in bovine terminal ileum and ileocecal valve
Beltramo C. Karymsakov T. Dondo A. Aryngaziyev B. Daugaliyeva A. Varello K. Acutis P.L. Daugaliyeva S. Peletto S.
2021Istituto Zooprofilattico dellAbruzzo e del Molise
Veterinaria Italiana
2021#57Issue 3209 - 214 pp.
The use of reference genes is commonly accepted as the most reliable approach to normalize qRT-PCR and to reduce possible errors in the quantification of gene expression. The aim of this study was to identify a set of reference genes suitable for gene expression analysis in the distal portion of small intestine and ileocecal valve in cattle. These sites of intestine are of interest in veterinary science as they are the main sites of inflammation caused by Mycobacterium avium subsp. paratuberculosis, agent of paratuberculosis. We employed ten PCR assays for commonly used reference genes belonging to various functional classes and then determined their expression stability. The most stable genes were RPL13A and HMBS, followed by TFRC and B-ACT. NormFinder analysis provided similar results with B-ACT as the best reference gene, followed by RPL13A and TFRC. This validated gene panel may be useful for studies on paratuberculosis aiming to identify genes differentially expressed by qRT-PCR.
Cattle , Normalization , Real-time qPCR , Reference genes , Small intestine
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Istituto Zooprofilattico Sperimentale del Piemonte, Ligura e Valle d’Aosta, via Bologna 148, Turin, 10154, Italy
LLP ‘Kazakh Research Institute for Livestock and Fodder Production’, st. Zhandosova 51, Almaty, Kazakhstan
LLP ‘Scientific Production Center of Microbiology and Virology’, Bogenbai Batyr str., 105, Almaty, Kazakhstan
Istituto Zooprofilattico Sperimentale del Piemonte
LLP ‘Kazakh Research Institute for Livestock and Fodder Production’
LLP ‘Scientific Production Center of Microbiology and Virology’
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