Evaluation of housekeeping genes as references for quantitative real time RT-PCR analysis of gene expression in Japanese flounder (Paralichthys olivaceus)

doi:10.1016/j.fsi.2010.12.014

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	Evaluation of housekeeping genes as references for quantitative real time RT-PCR analysis of gene expression in Japanese flounder (Paralichthys olivaceus)
	Zheng, Wen-jiang 1,2; Sun, Li 1
刊名	FISH & SHELLFISH IMMUNOLOGY
	2011-02-01
卷号	30 期号:2 页码:638-645
关键词	Quantitative Real Time Rt-pcr Reference Gene Housekeeping Gene Paralichthys Olivaceus
ISSN号	1050-4648
DOI	10.1016/j.fsi.2010.12.014
文献子类	Article
英文摘要	Japanese flounder (Paralichthys olivaceus) is an important economic fish species cultured worldwide. In this report, we compared the potentials of ten housekeeping genes as quantitative real time RT-PCR (qRT-PCR) references for the study of gene expression in Japanese flounder under normal physiological conditions and during bacterial infection. For this purpose, the expression of the ten genes in eight flounder tissues (liver, spleen, kidney, heart, muscle, brain, gill, and intestine) was determined by qRT-PCR before and after bacterial infection. The expression levels of the housekeeping genes were then compared and evaluated with geNorm and NormFinder algorithms. The results showed that before bacterial infection, the tested genes exhibited tissue-specific expressions to various degrees, with beta-actin and ubiquitin-conjugating enzyme being ranked as the most stable genes across tissue types. Following bacterial challenge, all the tested genes varied in expression levels in tissue-dependent manners and no cross-all-tissue type reference gene was identified among the examined panel of housekeeping genes; however, alpha-tubulin was recognized as the most stable gene in four (spleen, heart, muscle, and gill) of the eight examined tissues. These results indicate that for qRT-PCR analysis of gene expression in Japanese flounder as a function of bacterial infection, the choice of reference genes should be made according to tissue type. (C) 2010 Elsevier Ltd. All rights reserved.; Japanese flounder (Paralichthys olivaceus) is an important economic fish species cultured worldwide. In this report, we compared the potentials of ten housekeeping genes as quantitative real time RT-PCR (qRT-PCR) references for the study of gene expression in Japanese flounder under normal physiological conditions and during bacterial infection. For this purpose, the expression of the ten genes in eight flounder tissues (liver, spleen, kidney, heart, muscle, brain, gill, and intestine) was determined by qRT-PCR before and after bacterial infection. The expression levels of the housekeeping genes were then compared and evaluated with geNorm and NormFinder algorithms. The results showed that before bacterial infection, the tested genes exhibited tissue-specific expressions to various degrees, with beta-actin and ubiquitin-conjugating enzyme being ranked as the most stable genes across tissue types. Following bacterial challenge, all the tested genes varied in expression levels in tissue-dependent manners and no cross-all-tissue type reference gene was identified among the examined panel of housekeeping genes; however, alpha-tubulin was recognized as the most stable gene in four (spleen, heart, muscle, and gill) of the eight examined tissues. These results indicate that for qRT-PCR analysis of gene expression in Japanese flounder as a function of bacterial infection, the choice of reference genes should be made according to tissue type. (C) 2010 Elsevier Ltd. All rights reserved.
学科主题	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
URL标识	查看原文
语种	英语
WOS记录号	WOS:000288305300024
公开日期	2012-07-03
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/11858]
专题	海洋研究所_实验海洋生物学重点实验室
作者单位	1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 260071, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Zheng, Wen-jiang,Sun, Li. Evaluation of housekeeping genes as references for quantitative real time RT-PCR analysis of gene expression in Japanese flounder (Paralichthys olivaceus)[J]. FISH & SHELLFISH IMMUNOLOGY,2011,30(2):638-645.
APA	Zheng, Wen-jiang,&Sun, Li.(2011).Evaluation of housekeeping genes as references for quantitative real time RT-PCR analysis of gene expression in Japanese flounder (Paralichthys olivaceus).FISH & SHELLFISH IMMUNOLOGY,30(2),638-645.
MLA	Zheng, Wen-jiang,et al."Evaluation of housekeeping genes as references for quantitative real time RT-PCR analysis of gene expression in Japanese flounder (Paralichthys olivaceus)".FISH & SHELLFISH IMMUNOLOGY 30.2(2011):638-645.