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翘嘴鳜per1 mRNA表达量分析中采用的内参基因稳定性比较

Comparative Research on the Stability of Reference Genes Based on mRNA Expression Analysis of per1 in Siniperca chuatsi

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摘要

为筛选生物钟核心基因per1表达定量中的相对稳定性最好的内参基因, 本研究取翘嘴鳜成鱼心脏、肝脏、肾脏、脑、红肌、白肌、肠、眼和脾等九个组织为研究对象, 选取GAPDH、18S rRNA、β-actin、rps29、RPL13a、B2M和EF1a为内参基因, 采用实时荧光定量PCR(qRT-PCR)对per1基因mRNA表达水平进行检测分析。研究结果表明18S rRNA和GAPDH的平均稳定值M最低, 相对表达量最稳定。以18S rRNA和GAPDH为内参基因时分析发现per1基因表达量在肝脏中最高。本研究为在鱼类per1 mRNA表达检测过程中选用稳定的内参基因提供了实验和理论参考。

Abstract

For the screening the best reference gene of the biological clock core gene per1 expression in quantitative analysis, we compared the stability of the 7 reference genes (GAPDH, 18S rRNA, β-actin, rps29, RPL13a, B2M and EF1a) in heart, brain, kidney, liver, slow muscle, fast muscle, intestine, spleen and eye tissues of the adult Siniperca chuatsi. By using GeNorm software, the results showed that the GAPDH and 18S rRNA are the the most stable reference genes in 9 tissues. With 18S rRNA and GAPDH as reference gene analysis of the expression of per1 in the liver was the highest. This study for the detection of per1 mRNA levels in fish tissues and in the selection of one or more reference genes provide experimental and theoretical reference.

Newport宣传-MKS新实验室计划
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中图分类号:Q786

DOI:10.3969/j.issn.1007-7146.2017.04.015

所属栏目:基础研究

基金项目:湖南省教育厅重点实验室创新平台开放基金(15K011); 长沙市科技计划重点项目(ZD1601003); 国家自然科学基金(31640086)

收稿日期:2017-05-02

修改稿日期:2017-05-12

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吴 萍:长沙学院生物与环境工程学院, 湖南 长沙410003湖南省水产高效健康生产协同创新中心, 湖南 常德 415000
张方亮:长沙学院生物与环境工程学院, 湖南 长沙410003湖南省水产高效健康生产协同创新中心, 湖南 常德 415000
杨程涌:长沙学院生物与环境工程学院, 湖南 长沙410003湖南省水产高效健康生产协同创新中心, 湖南 常德 415000
张建社:长沙学院生物与环境工程学院, 湖南 长沙410003湖南省水产高效健康生产协同创新中心, 湖南 常德 415000
褚武英:长沙学院生物与环境工程学院, 湖南 长沙410003湖南省水产高效健康生产协同创新中心, 湖南 常德 415000

联系人作者:吴萍(249458368@qq.com)

备注:吴萍(1990-), 女, 湖南长沙人, 博士研究生, 研究方向基因调控。

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引用该论文

WU Ping,ZHANG Fangliang,YANG Chengyong,ZHANG Jianshe,CHU Wuying. Comparative Research on the Stability of Reference Genes Based on mRNA Expression Analysis of per1 in Siniperca chuatsi[J]. Acta Laser Biology Sinica, 2017, 26(4): 379-384

吴 萍,张方亮,杨程涌,张建社,褚武英. 翘嘴鳜per1 mRNA表达量分析中采用的内参基因稳定性比较[J]. 激光生物学报, 2017, 26(4): 379-384

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