A new set of endogenous reference genes for gene expression studies of porcine stomach

Abstract

Gene expression analysis by Real-Time PCR requires careful selection of endogenous reference genes to obtain accurate results. We evaluated usefulness of six reference genes to expression studies of porcine stomach. We selected genes which were recently described as very stable (RPL27, RPS29, RPS13, OAZ1) and two commonly used housekeeping genes (GAPDH, ACTB). Our results indicate that OAZ1, RPS29 and RPL27 are more suitable reference genes than ACTB and GAPDH in expression studies of porcine stomach. In our study the most stable genes were OAZ1 (M=0.856), RPS29 (M=0.862) and RPL27 (M=0.892), while ACTB (M=1.03) and GAPDH (M=1.005) were less stable. RPS13 (M=1.913) appeared to be highly unstable. Set of three the most stable genes has been used to compare ghrelin (GHRL) expression in two different regions (diverticulum ventriculi and fundus ventriculi) of porcine stomach. Ghrelin mRNA was highly expressed in both regions of stomach, however level of GHRL mRNA was approximately 5-fold higher in fundus ventriculi than in diverticulum ventriculi.