Chen X J, Zhang X Q, Huang S, Cao Z J, Qin Q W, Hu W T, Sun Y, Zhou Y C
University, P.R.
, P. R.
Pol J Vet Sci. 2017 Sep 26;20(3):583-594. doi: 10.1515/pjvs-2017-0071.
Golden pompano (Trachinotus ovatus) is an important economically fish species. In this study, with an aim to identify reliable reference genes for quantitative real-time PCR (qRT-PCR) in golden pompano, we evaluated the expression stability of eight housekeeping genes in the presence and absence of poly I:C stimulation in eight tissues. The PCR data was analyzed by geNorm and NormFinder algorithms. The results showed that the expression of all the examined genes exhibited tissue-dependent variations. When under normal physiological condition, geNorm and NormFinder identified B2M and 18S as suitable genes. When studying gene expression under conditions of poly I:C stimulation, the selection of the internal controls should be selected on a tissue basis. At 12 h stimulation, geNorm ranked Actin/UBCE, Actin/B2M, UBCE/B2M, Actin/UBCE, RPL13/B2M, UBCE/GAPDH, B2M/RPL13, and UBCE/B2M, respectively, as the most stably expressed genes in liver, spleen, kidney, gill, intestine, heart, muscle, and brain. Comparable ranking orders were produced by NormFinder. Similar results were obtained at 48 h stimulation. Taken together, these results indicate that B2M and 18S are the most stable gene across tissue types under normal physiological conditions. However, during poly I:C stimulation, no single gene or single pair of genes in the examined set of housekeeping genes can serve as a universal reference across all tissue types. If one gene is preferred, B2M, B2M, UBCE, Actin, B2M/RPL13, B2M, B2M, and RPL13 may be used in spleen, kidney, liver, gill, intestine, brain, muscle, and heart of golden pompano, respectively.
金钱鱼（Trachinotus ovatus）是一种重要的经济鱼类。在本研究中，为了鉴定金钱鱼定量实时PCR（qRT-PCR）中可靠的内参基因，我们评估了8个管家基因在8种组织中有无聚肌胞苷酸（poly I:C）刺激情况下的表达稳定性。通过geNorm和NormFinder算法分析PCR数据。结果表明，所有检测基因的表达均呈现组织依赖性变化。在正常生理条件下，geNorm和NormFinder鉴定B2M和18S为合适的基因。在研究poly I:C刺激条件下的基因表达时，应根据组织选择内参。在刺激12小时时，geNorm分别将肌动蛋白/泛素羧基末端水解酶（Actin/UBCE）、肌动蛋白/B2M、UBCE/B2M、肌动蛋白/UBCE、核糖体蛋白L13（RPL13）/B2M、UBCE/甘油醛-3-磷酸脱氢酶（GAPDH）、B2M/RPL13和UBCE/B2M列为肝脏、脾脏、肾脏、鳃、肠道、心脏、肌肉和大脑中表达最稳定的基因。NormFinder得出了类似的排序结果。在刺激48小时时也获得了类似结果。综上所述，这些结果表明，在正常生理条件下，B2M和18S是所有组织类型中最稳定的基因。然而，在poly I:C刺激期间，所检测的管家基因集中没有单个基因或单对基因可作为所有组织类型的通用内参。如果首选一个基因，B2M、B2M、UBCE、肌动蛋白、B2M/RPL13、B2M、B2M和RPL13可分别用于金钱鱼的脾脏、肾脏、肝脏、鳃、肠道、大脑、肌肉和心脏。
