title: Evaluation of potential reference genes for relative quantification by RT-qPCR in different porcine tissues derived from feeding studies.
abstract: Five potential reference genes for RT-qPCR application, namely histone H3, beta-actin, GAPDH, ubiquitin and 18S rRNA, were evaluated for normalization of gene expression in four selected tissues (liver, kidney, thyroid and abdominal fat). Tissues were derived from fattening pigs exposed to different amounts and type of dietary iodine. Two software applications (geNorm and NormFinder) were used to evaluate the stability of the potential reference genes. All studied genes displayed high expression stability but different stability patterns between the investigated tissues. The results suggest GAPDH and 18S rRNA as reference genes applicable in all tissues investigated. Beta-actin and histone H3 are suitable reference genes for all tissues investigated except fat. In contrast, ubiquitin should be excluded from use as a reference gene in the porcine tissues analyzed due to variations in expression levels, despite the good expression stability.
authors.affiliation: Institute of Animal Nutrition, Products, and Nutrition Physiology (APN), University of Natural Resources and Life Sciences, 1190 Vienna, Austria; E-Mails: thomas.ettle@lfl.bayern.de (T.E); christiane.mair@boku.ac.at (C.M.); qimeng.li@boku.ac.at (Q.L.).
authors.forename: Qimeng
authors.initials: Q
authors.lastname: Li
authors.forename: Konrad Johann
authors.initials: KJ
authors.lastname: Domig
authors.forename: Thomas
authors.initials: T
authors.lastname: Ettle
authors.forename: Wilhelm
authors.initials: W
authors.lastname: Windisch
authors.forename: Christiane
authors.initials: C
authors.lastname: Mair
authors.forename: Karl
authors.initials: K
authors.lastname: Schedle
mesh_terms: D050153:Abdominal Fat / Q000378:metabolism; D000199:Actins / Q000235:genetics; D000818:Animals; D015870:Gene Expression; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000235:genetics; D006657:Histones / Q000235:genetics; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D012313:RNA / Q000378:metabolism / Q000592:standards*; D020871:RNA Stability; D012335:RNA, Ribosomal / Q000235:genetics; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards*; D013552:Swine; D013961:Thyroid Gland / Q000378:metabolism; D025801:Ubiquitin / Q000235:genetics
chemical_list: D000199:Actins; D006657:Histones; D012335:RNA, Ribosomal; D025801:Ubiquitin; D012313:RNA; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)