Postnatal jejunal expression patterns of four major house‐keeping genes in pigs are measured by the real time RT‐PCR

Abstract

The selection of an ideal house-keeping gene is essential to examine developmental changes in the intestinal gene expression studies by real time RT-PCR. This study was carried out to examine the postnatal porcine jejunal expression patterns of four major house-keeping genes, beta-actin (ACTIN), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyl-transferase (HPRT) and ribosomal protein L10 (RPL10) by real-time RT-PCR. Thirty six purebred Yorkshire gilts were used for sampling jejunum at the ages of d 1, 4, 6, 12, 20 and 28 (1 wk post-weaning). The house-keeping gene expression, indirectly expressed as threshold cycles (CT) values, was analyzed by real time RT-PCR (SmartCycler) using the SYBR Green-I detection kit. Orthogonal polynomial contrasts showed no changes (P>0.05) in ACTIN (16.1–16.8; SE=0.56), GAPDH (19.4–20.6; SE=0.50) and HPRT (28.0–29.8; SE=0.98) expressions but quartic responses in RPL10 (P=0.006; 17.7–18.9, SE=0.25) expression. Our results indicate that beta-actin, GAPDH and HPRT are the ideal housekeeping genes to examine porcine jejunal gene ontogeny research. Supported by NSERC of Canada and the Chinese Academy of Sciences.