利用RT-qPCR技术筛选菊黄东方鲀的最适内参基因  

Selection of appropriate reference genes for real-time quantitative PCR analysis in Takifugu flavidus

作　　者：蒋彩云 乔琨[2] 许旻[2] 熊静[1] 刘智禹[2,3] 黄文树 JIANG Caiyun;QIAO Kun;XU Min;XIONG Jing;LIU Zhiyu;HUANG Wenshu(Fisheries College of Jimei University,Xiamen 361021,China;Key Laboratory of Cultivation and High-value Utilization of Marine Organisms in Fujian Province,Fisheries Research Institute of Fujian,Xiamen 361013,China;Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources,Xiamen 361021,China)

机构地区：[1]集美大学水产学院,福建厦门361021 [2]福建省水产研究所,福建省海洋物增养殖与高值化利用重点实验室,福建厦门361013 [3]福建省海洋生物资源开发利用协同创新中心,福建厦门361021

出　　处：《渔业研究》2020年第2期105-116,共12页Journal of Fisheries Research

基　　金：福建省种业创新与产业化工程项目(2017FJSCZY03);福建省海洋渔业结构调整专项(2017HYJG07、2017HYJG08);2019年度福建省海洋经济发展专项.

摘　　要：定量PCR技术已经成为基因表达水平测定最常用的方法,选择合适的内参基因对基因表达水平的准确定量至关重要。本文检测了菊黄东方鲀β-actin、GAPDH、EF1-α和18S rRNA等4种内参基因在不同组织、生长环境和发育阶段中的表达,应用3种内参基因筛选软件(geNorm、NormFinder和Bestkeeper)综合分析了这4种内参基因表达的稳定性。结果表明内参基因在成鱼养殖和野生菊黄东方鲀内的稳定性排名均为:EF1-α>β-actin>18S rRNA>GAPDH;在养殖菊黄东方鲀3月龄、6月龄和12月龄三个发育阶段中内参基因EF1-α最稳定,其他各组织在不同发育阶段内参基因的选择有所差异。研究结果为不同条件下菊黄东方鲀功能基因表达的定量分析提供重要参考。Quantitative PCR technology has become the most commonly used method for the gene expression level determination.Choosing the appropriate reference gene is very important for the accurate quantitative gene to gene expression level.In this paper,the expressions of four internal reference genes,includingβ-actin、GAPDH、EF1-αand 18S,in different tissues,growth environments and development stages of T.flavidus were detected,and the stability of the expression of these four internal reference genes was comprehensively analyzed by using three internal reference gene screening software(geNorm,NormFinder and Bestkeeper).The results showed that the stable rank of the internal reference gene in adult and wild puffer fish was EF1-α>β-actin>18S>GAPDH.Among the three developmental stages of puffer fish(3 months,6 months and 12 months),EF1-αgene was the most stable,while other tissues had different selection of reference genes in different development stages.The results provided an important reference for quantitative analysis of functional gene expression in T.flavidus under different conditions.

关 键 词：菊黄东方鲀 RT-QPCR 内参基因 geNorm软件 

分 类 号：Q344.13[生物学—遗传学] Q781