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Evaluation of potential internal references for quantitative real-time RT-PCR normalization of gene expression in red drum (Sciaenops ocellatus)
Sun, Bo-guang; Hu, Yong-hua
2015-06-01
发表期刊FISH PHYSIOLOGY AND BIOCHEMISTRY
卷号41期号:3页码:695-704
文章类型Article
摘要Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) has been used extensively for studying gene expression in diverse organisms including fish. In this study, with an aim to identify reliable reference genes for qRT-PCR in red drum (Sciaenops ocellatus), an economic fish species, we determined the expression stability of seven housekeeping genes in healthy and bacterium-infected red drum. Each of the selected candidate genes was amplified by qRT-PCR from the brain, gill, heart, intestine, kidney, liver, muscle, and spleen of red drum infected with or without a bacterial pathogen for 12 and 48 h. The mRNA levels of the genes were analyzed with the geNorm and NormFinder algorithms. The results showed that in the absence of bacterial infection, translation initiation factor 3, NADH dehydrogenase 1, and QM-like protein may be used together as internal references across the eight examined tissues. Bacterial infection caused variations in the rankings of the most stable genes in a tissue-dependent manner. For all tissues, two genes sufficed for reliable normalization at both 12 and 48 h post-infection. However, the optimal gene pairs differed among tissues and, for four of the examined eight tissues, between infection points. These results indicate that when studying gene expression in red drum under conditions of bacterial infection, the optimal reference genes should be selected on the basis of tissue type and, for accurate normalization, infection stage.
关键词Housekeeping Gene Expression Stability Reference Gene Sciaenops Ocellatus
DOI10.1007/s10695-015-0039-8
收录类别SCI
语种英语
WOS记录号WOS:000354619300011
引用统计
被引频次:12[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/26965
专题实验海洋生物学重点实验室
作者单位Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China
第一作者单位实验海洋生物学重点实验室
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Sun, Bo-guang,Hu, Yong-hua. Evaluation of potential internal references for quantitative real-time RT-PCR normalization of gene expression in red drum (Sciaenops ocellatus)[J]. FISH PHYSIOLOGY AND BIOCHEMISTRY,2015,41(3):695-704.
APA Sun, Bo-guang,&Hu, Yong-hua.(2015).Evaluation of potential internal references for quantitative real-time RT-PCR normalization of gene expression in red drum (Sciaenops ocellatus).FISH PHYSIOLOGY AND BIOCHEMISTRY,41(3),695-704.
MLA Sun, Bo-guang,et al."Evaluation of potential internal references for quantitative real-time RT-PCR normalization of gene expression in red drum (Sciaenops ocellatus)".FISH PHYSIOLOGY AND BIOCHEMISTRY 41.3(2015):695-704.
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