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Evaluation of reference genes for quantitative real-time RT-PCR analysis of gene expression in Nile tilapia (Oreochromis niloticus)

文献类型： 外文期刊

作者： Yang, Chang Geng 1 ; Wang, Xian Li 2 ; Tian, Juan 1 ; Liu, Wei 1 ; Wu, Fan 1 ; Jiang, Ming 1 ; Wen, Hua 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Key Lab Freshwater Biodivers Conservat & Utilizat, Minist Agr, Yangtze River Fisheries Res Inst, Wuhan 430223, Peoples R China

2.Tongji Univ, Sch Med, Sarite Ctr Stem Cell Engn Translat Med, East Hosp,Stem Cell Res Ctr, Shanghai

关键词： Reference genes;RT-qPCR;Tilapia

期刊名称：GENE （ 影响因子：3.688； 五年影响因子：3.329 ）

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收录情况： SCI

摘要： Quantitative real-time reverse-transcriptase polymerase chain reaction (RT-qPCR) has been used frequently to study gene expression related to fish immunology. In such studies, a stable reference gene should be selected to correct the expression of the target gene. In this study, seven candidate reference genes (glyceraldehyde-3-phosphate dehydrogenase (GADPH), ubiquitin-conjugating enzyme (UBCE), 18S ribosomal RNA (18S rRNA), beta-2-microglobulin (B2M), elongation factor 1 alpha (EF1A), tubulin alpha chain-like (TUBA) and beta actin (ACTB)), were selected to analyze their stability and normalization in seven tissues (liver, spleen, kidney, brain, heart, muscle and intestine) of Nile tilapia (Oreochromis niloticus) challenged with Streptococcus agalactiae or Streptococcus iniae, respectively. The results showed that all the candidate reference genes exhibited tissue-dependent transcriptional variations. With PBS injection as a control, UBCE was the most stable and suitable single reference gene in the intestine, liver, brain, kidney, and spleen after S. iniae infection, and in the liver, kidney, and spleen after S. agalactiae infection. EF1A was the most suitable in heart and muscle after S. iniae or S. agalactiae infection. GADPH was the most suitable gene in intestine and brain after S. agalactiae infection. In normal conditions, UBCE and 18S rRNA were the most stably expressed genes across the various tissues. These results showed that for RT-qPCR analysis of tilapia, selecting two or more reference genes may be more suitable for cross-tissue analysis of gene expression.

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