斑鳢实时定量PCR内参基因的分离鉴定
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- 英文论文题名：Identification of suitable reference genes for quantitative real-time PCR(qRT-PCR) normalization in Blotched Snakehead(Channa maculate)
- 论文作者：茅华华 ; 陈昆慈 ; 赵建 ; 罗青 ; 徐红艳
- 英文论文作者：MAO Huahua ; CHEN Kunci ; ZHAO Jian ; LUO Qing ; XU Hongyan ; Pearl River Fisheries Research Institute ; Chinese Academy of Fishery Sciences ; College of Fisheries and Life Science ; Shanghai Ocean University
- 年：2016
- 作者机构：中国水产科学研究院珠江水产研究所农业部热带亚热带水产资源利用与养殖重点实验室;上海海洋大学水产与生命学院;
- 论文关键词：内参基因 ; qRT-PCR ; 基因克隆 ; 斑鳢
- 英文论文关键词：Reference genes ; qRT-PCR ; Gene cloning ; Blotched Snakehead
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S917.4
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：573k
- 原文格式：D
- 会议级别：全国
摘要
斑鳢是我国重要的经济鱼类之一。为了应用荧光定量PCR技术准确地分析斑鳢功能基因的时序及组织特异性表达,本研究克隆分析了9个基因斑鳢的候选内参基因,包括:actb,tmem104,ef1α,tmem256,sep15等。首先通过实时荧光定量PCR技术,检测了这9个内参基因在成鱼不同组织(心、肝、脾、肾、脑、眼、性腺和肌肉)和胚胎不同发育时期的mRNA表达情况。然后通过Ref Find软件对这些基因的表达稳定性进行了分析和评估,结果表明:actin-β和tmem104在成鱼不同组织表达最稳定,sep15和tmem256在胚胎不同发育时期的表达最稳定;因此,actin-β和tmem104适合作为斑鳢基因组织表达分析的内参基因,而sep15和tmem256可为胚胎发育时期的内参基因。
The blotched snakehead is one of the most important economic fish in China. To accurately analyze the gene expression in this fish, the stable and reliable reference genes are required for the normalization of real-time quantitative PCR(qRT-PCR). Here, 9 candidate genes were isolated from blotched snakehead, including actb, tmem104, ef1α, tmem256, sep15, and so on. Their expression levels were examined in different development stages of embryos and different tissue types(heart, liver, spleen, kidney, brain, eye, gonads and muscle) by RT-qPCR.The genes expression stability were assessed using Ref Find, the results showed, actin-β and tmem104 were the most stable two for different tissue types, and sep15 and tmem256 were the most stable genes for stages of embryos. That is to say, these stable genes could be used as reliable reference genes for normalization of qRT-PCR data in the blotched snakehead.
The blotched snakehead is one of the most important economic fish in China. To accurately analyze the gene expression in this fish, the stable and reliable reference genes are required for the normalization of real-time quantitative PCR(qRT-PCR). Here, 9 candidate genes were isolated from blotched snakehead, including actb, tmem104, ef1α, tmem256, sep15, and so on. Their expression levels were examined in different development stages of embryos and different tissue types(heart, liver, spleen, kidney, brain, eye, gonads and muscle) by RT-qPCR.The genes expression stability were assessed using Ref Find, the results showed, actin-β and tmem104 were the most stable two for different tissue types, and sep15 and tmem256 were the most stable genes for stages of embryos. That is to say, these stable genes could be used as reliable reference genes for normalization of qRT-PCR data in the blotched snakehead.
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