Shui Fei, Qiu Guiru, Pan Shenqiang, Wang Xin, Jia Fumin, Jiang Tingting, Li Yongsheng, Geng Zhaoyu, Jin Sihua
College of Animal Science and Technology, Anhui Agricultural University, Hefei, China.
Anhui Provincial Key Laboratory of Local Animal Genetic Resources Conservation and Bio-Breeding, Hefei, China.
Front Vet Sci. 2023 Apr 3;10:1160384. doi: 10.3389/fvets.2023.1160384. eCollection 2023.
Quantitative polymerase chain reaction (qPCR) is an important method to detect gene expression at the molecular level. The selection of appropriate housekeeping genes is the key to accurately calculating the expression level of target genes and conducting gene function studies. In this study, the expression of eight candidate reference genes, glyceraldehyde-3-phosphate dehydrogenase (), beta-actin (β), 18S ribosomal RNA (), hydroxymethylbilane synthase (), hypoxanthine phosphoribosyltransferase 1 (), TATA box binding protein (), ribosomal protein L13 (), and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein (), in the duodenal epithelial tissue of 42-day-old meat-type ducks were detected using qPCR. Furthermore, their expression stability was analyzed using the geNorm, NormFinder, and BestKeeper programs. The results indicated that and were the most stably expressed genes. All three programs indicated that the expression of was the least stable, making it unsuitable for the study of gene expression in meat-type duck tissues. This study provides stable reference genes for gene expression analysis and contributes to further studies on the gene function of meat-type ducks.
定量聚合酶链反应（qPCR）是在分子水平检测基因表达的重要方法。选择合适的管家基因是准确计算靶基因表达水平和进行基因功能研究的关键。在本研究中，使用qPCR检测了42日龄肉鸭十二指肠上皮组织中8个候选内参基因的表达，这些基因分别是甘油醛-3-磷酸脱氢酶（）、β-肌动蛋白（β）、18S核糖体RNA（）、羟甲基胆色素原合酶（）、次黄嘌呤磷酸核糖基转移酶1（）、TATA盒结合蛋白（）、核糖体蛋白L13（）和酪氨酸3-单加氧酶/色氨酸5-单加氧酶激活蛋白（）。此外，使用geNorm、NormFinder和BestKeeper程序分析了它们的表达稳定性。结果表明，和是表达最稳定的基因。所有三个程序均表明的表达最不稳定，不适用于肉鸭组织基因表达的研究。本研究为基因表达分析提供了稳定的内参基因，有助于肉鸭基因功能的进一步研究。
