Azarpeykan Sara, Dittmer Keren E
Institute of Veterinary, Animal and Biomedical Science, Massey University, Palmerston North 4442, New Zealand.
J Equine Sci. 2016;27(4):165-168. doi: 10.1294/jes.27.165. Epub 2016 Dec 15.
Housekeeping genes (HKGs) are used as internal controls for normalising and calculating the relative expression of target genes in RT-qPCR experiments. There is no unique universal HKG and HKGs vary among organisms and tissues, so this study aimed to determine the most stably expressed HKGs in the equine kidney. The evaluated HKGs included 18S ribosomal RNA (18S), 28S ribosomal RNA (28S), ribosomal protein L32 (RPL32), β-2-microglobulin (B2M), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), succinate dehydrogenase complex (SDHA), zeta polypeptide (YWHAZ), and hypoxanthine phosphoribosyltransferase 1 (HPRT1). The HKGs expression stability data were analysed with two software packages, geNorm and NormFinder. The lowest stability values for geNorm suggests that YWHAZ and HPRT1 would be most optimal (M=0.31 and 0.32, respectively). Further, these two genes had the best pairwise stability value using NormFinder (geNorm V=0.085). Therefore, these two genes were considered the most useful for RT-qPCR studies in equine kidney.
管家基因（HKGs）在逆转录定量聚合酶链反应（RT-qPCR）实验中用作内参，用于标准化和计算靶基因的相对表达量。不存在唯一通用的管家基因，不同生物体和组织中的管家基因各不相同，因此本研究旨在确定马肾中表达最稳定的管家基因。评估的管家基因包括18S核糖体RNA（18S）、28S核糖体RNA（28S）、核糖体蛋白L32（RPL32）、β-2-微球蛋白（B2M）、甘油醛-3-磷酸脱氢酶（GAPDH）、琥珀酸脱氢酶复合物（SDHA）、ζ多肽（YWHAZ）和次黄嘌呤磷酸核糖基转移酶1（HPRT1）。使用geNorm和NormFinder这两个软件包分析管家基因的表达稳定性数据。geNorm的最低稳定性值表明YWHAZ和HPRT1最适合（M值分别为0.31和0.32）。此外，使用NormFinder时，这两个基因的成对稳定性值最佳（geNorm V = 0.085）。因此，这两个基因被认为对马肾的RT-qPCR研究最有用。
