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Quantitative real time RT-PCR study of pathogen-induced gene expression in rock bream (Oplegnathus fasciatus): Internal controls for data normalization
Zhang, Bao-Cun1,2; Sun, Li1; Xiao, Zhi-zhong1; Hu, Yong-hua1; Hu, YH (reprint author), Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China.
2014-06-01
发表期刊MARINE GENOMICS
卷号15页码:75-84
文章类型Article
摘要Rock bream Oplegnathus fasciatus is an important economic fish species. In this study, we evaluated the appropriateness of six housekeeping genes as internal controls for quantitative real-time PCR (RT-qPCR) analysis of gene expression in rock bream before and after pathogen infection. The expression of the selected genes in eight tissues infected with Vibrio alginolyticus or megalocytivirus was determined by RT-qPCR, and the PCR data were analyzed with geNorm and NormFinder algorithms. The results showed that before pathogen infection, mediator of RNA polymerase II transcription subunit 8 and beta-actin were ranked as the most stable genes across the examined tissues. After bacterial or viral infection, the stabilities of the housekeeping genes varied to significant extents in tissue-dependent manners, and no single pair of genes was identified as suitable references for all tissues for either of the pathogen stimuli. In addition, for the majority of tissues, the most stable genes during bacterial infection differed from those during viral infection. Nevertheless, optimum reference genes were identified for each tissue under different conditions. Taken together, these results indicate that tissue type and the nature of the infectious agent used in the study can all influence the choice of normalization factors, and that the optimum reference genes identified in this study will provide a useful guidance for the selection of internal controls in future RT-PCR study of gene expression in rock bream. (C) 2014 Elsevier B.V. All rights reserved.
关键词Quantitative Real Time Rt-pcr Internal Reference Housekeeping Gene Oplegnathus Fasciatus
学科领域Genetics & Heredity ; Marine & Freshwater Biology
DOI10.1016/j.margen.2014.03.001
收录类别SCI
语种英语
WOS记录号WOS:000338606500013
引用统计
被引频次:13[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/337002/24156
专题实验海洋生物学重点实验室
通讯作者Hu, YH (reprint author), Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China.
作者单位1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
第一作者单位中国科学院海洋研究所
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Zhang, Bao-Cun,Sun, Li,Xiao, Zhi-zhong,et al. Quantitative real time RT-PCR study of pathogen-induced gene expression in rock bream (Oplegnathus fasciatus): Internal controls for data normalization[J]. MARINE GENOMICS,2014,15:75-84.
APA Zhang, Bao-Cun,Sun, Li,Xiao, Zhi-zhong,Hu, Yong-hua,&Hu, YH .(2014).Quantitative real time RT-PCR study of pathogen-induced gene expression in rock bream (Oplegnathus fasciatus): Internal controls for data normalization.MARINE GENOMICS,15,75-84.
MLA Zhang, Bao-Cun,et al."Quantitative real time RT-PCR study of pathogen-induced gene expression in rock bream (Oplegnathus fasciatus): Internal controls for data normalization".MARINE GENOMICS 15(2014):75-84.
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