title: Selection of reference genes for studies of porcine endometrial gene expression on gestational day 12.
abstract: Comparing gene expression patterns in the endometrium on gestational day 12 (GD12) between Erhualian (ER) and Landrace×Large White (LL) pigs is helpful to understand the biological mechanisms of fecundity. Selecting genes that have stable expression levels as the internal standards in a comparative study is essential for identifying real gene-specific variation by quantitative RT-PCR (qRT-PCR). Five genes expressed in sow endometria on GD12 were evaluated for their suitability as internal control for relative quantification by qRT-PCR. These genes were beta-actin (ACTB), beta-2-microglobulin (B2M), phosphoglycerate kinase 1 (PGK1), RNA polymerase II polypeptide G (RPG), and ribosomal protein S20 (RPS20), which represent different functional classes. Our results indicated that ACTB, B2M, and PGK1 were not suitable as internal standards for normalization because of their huge variability between the two breeds. RPS20 and RPG were most stable, and the former is recommended to serve as the internal standard when the use of multiple housekeeping genes is unpractical.
authors.affiliation: Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou, Guangdong 510642, China. shouqiw@163.com
authors.forename: Shouqi
authors.initials: S
authors.lastname: Wang
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Li
authors.forename: Ailing
authors.initials: A
authors.lastname: Zhang
authors.forename: Manqing
authors.initials: M
authors.lastname: Liu
authors.forename: Hao
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000818:Animals; D004717:Endometrium / Q000378:metabolism*; D005260:Female; D005298:Fertility / Q000235:genetics*; D020869:Gene Expression Profiling / Q000592:standards*; D018507:Gene Expression Regulation, Developmental; D011247:Pregnancy; D012319:RNA Polymerase II / Q000235:genetics*; D012015:Reference Standards; D012269:Ribosomal Proteins / Q000235:genetics*; D034421:Sus scrofa / Q000235:genetics*
chemical_list: D012269:Ribosomal Proteins; C029611:ribosomal protein S20; D012319:RNA Polymerase II