草鱼定量实时 PCR 内参基因的验证。
Validation of reference genes of grass carp Ctenopharyngodon idellus for the normalization of quantitative real-time PCR.
机构信息
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of Tropical & Subtropical Fish Breeding & Cultivation, CAFS, Guangzhou 510380, People's Republic of China.
出版信息
Biotechnol Lett. 2010 Aug;32(8):1031-8. doi: 10.1007/s10529-010-0258-0. Epub 2010 Apr 2.
Expression of four reference genes of grass carp, including beta-actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S rRNA (18S) and elongation factor-1 alpha (EF1alpha), was studied in tissues of normal individuals and bacteria-infected individuals. EF1alpha had the most stable expressions followed by 18S rRNA then GAPDH; ACTB had the least stability. After being infected with bacteria, the grass carp showed minimal changes in expression levels of EF1alpha in the liver and head kidney, while ACTB had the most stable expressions in spleen but the least stable in liver. EF1alpha is thus the optimal reference gene in quantitative real-time PCR analysis to quantitate the expression levels of target genes in tissues of grass carp.
本研究旨在探讨草鱼 4 种内参基因（β-肌动蛋白 ACTB、甘油醛-3-磷酸脱氢酶 GAPDH、18S 核糖体 RNA 18S 和延伸因子 1α EF1α）在正常个体和细菌感染个体组织中的表达情况。结果显示，EF1α 的表达最稳定，其次是 18S rRNA，然后是 GAPDH；而 ACTB 的表达最不稳定。在细菌感染后，草鱼肝脏和头肾中 EF1α 的表达水平变化最小，而 EF1α 在脾脏中表达最稳定，在肝脏中表达最不稳定。因此，EF1α 是草鱼组织定量 real-time PCR 分析中定量目标基因表达水平的最佳内参基因。
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