Stability of Reference Gene Expression After Porcine Sapelovirus Infection in Porcine Intestinal Epithelial Cells.
Intestinal epithelial cells, which serve as the first physical barrier to protect intestinal tract from external antigens, have an important role in the local innate immunity. Screening of reference genes that have stable expression levels after viral infection in porcine intestinal epithelial cells is critical for ensuring the reliability of the expression analysis on anti-infection genes in porcine intestinal epithelial cells. In this study, nine common reference genes in pigs, including ACTB, B2M, GAPDH, HMBS, SDHA, HPRT1, TBP, YWHAZ, and RPL32, were chosen as the candidate reference genes. Porcine sapelovirus (PSV) was used as a model virus to infect porcine intestinal epithelial cell line (IPEC-J2). The expression stability of the nine genes was assessed by the geNorm, NormFinder, and BestKeeper software. Moreover, RefFinder program was used to evaluate the analytical results of above three softwares, and a relative expression experiment of selected target gene was used to verify the analysis results. The comprehensive results indicated that the gene combination of TBP and RPL32 has the most stable expression, which could be considered as an appropriate reference gene for research on gene expression after PSV infection in IPEC-J2cells. The results provided essential data for expression analysis of anti-infection genes in porcine intestinal epithelial cells.
1 College of Life Science and Technology, Southwest University for Nationalities , Chengdu, China .
Yong
Y
Huang
1 College of Life Science and Technology, Southwest University for Nationalities , Chengdu, China .
Yabing
Y
Chen
2 Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University , Shanghai, China .
Huan
H
Sun
1 College of Life Science and Technology, Southwest University for Nationalities , Chengdu, China .|3 Institute of Qinghai-Tibetan Plateau, Southwest University for Nationalities , Chengdu, China .
Daoliang
D
Lan
D000818:Animals; D002460:Cell Line; D004847:Epithelial Cells / Q000821:virology*; D020869:Gene Expression Profiling / Q000592:standards*; D054884:Host-Pathogen Interactions; D007413:Intestinal Mucosa / Q000473:pathology / Q000821:virology; D010849:Picornaviridae / Q000254:growth & development*; D012015:Reference Standards; D013552:Swine