白羽王鸽不同组织RT-qPCR内参基因的筛选  

Screening of Reference Genes for RT-qPCR in Different Tissues of White King Pigeon

作　　者：陈兰 郭丽燕[1,2] 张涛 张闪闪[1,2] 顾若君[3] 张跟喜 谢恺舟[1,2] 王金玉 CHEN Lan;GUO Liyan;ZHANG Tao;ZHANG Shanshan;GU Ruojun;ZHANG Genxi;XIE Kaizhou;WANG Jinyu(College of Animal Science and Technology,Yangzhou University,Jiangsu Yangzhou 225009,China;International Cooperation Laboratory of Agriculture and Agricultural Products Safty,Jiangsu Yanghzou 225009,China;SanXiangAn Ecological agriculture Technology CO.LTD,Jiangsu Wuxi 214100,China)

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009 [2]教育部农业与农产品安全国际合作联合实验室,江苏扬州225009 [3]无锡三乡岸农业生态农业科技有限公司,江苏无锡214100

出　　处：《中国畜牧杂志》2020年第1期77-82,共6页Chinese Journal of Animal Science

基　　金：江苏高校优势学科建设工程

摘　　要：内参基因的正确选择是获得精准、可靠RT-qPCR数据的重要前提。本研究随机选取3只28日龄的白羽王鸽,采集心、肝、脾、肺、肾、胸肌、腿肌、腹脂、卵巢9个组织作为实验材料,选取GAPDH、β-actin、18S rRNA和RPS24个常见的内参基因作为候选内参基因。利用RT-qPCR技术和geNorm、NormFinder、BestKeeper软件及Ct值分析法对候选基因在不同组织中的表达进行检测和稳定性分析。结果表明:geNorm、NormFinder软件分析与Ct值分析法的结果一致,即18S rRNA基因稳定性最佳,RPS2次之;BestKeeper软件的分析结果显示RPS2基因表达稳定性最好,18S rRNA基因次之。因此,推荐选择RPS2+18S rRNA的内参基因组合用于白羽王鸽不同组织基因表达的研究。The correct choice of the internal reference gene is an important prerequisite for obtaining accurate and reliable RT-qPCR data.In this study,heart,liver,spleen,lung,kidney,breast muscle,leg muscle,abdominal fat and ovary tissues were collected from three 28-day-old white king pigeons.Four common reference genes of GAPDH,β-actin,18S rRNA and RPS2 were selected as candidate internal reference genes.The expression and stability of the four candidate reference genes in different tissues were detected and analyzed by RT-qPCR and geNorm,NormFinder,BestKeeper software and Ct value analysis.It was found that the results analyzed using NormFinder software are consistent with that analyzed using Ct value analysis method.The 18S rRNA gene has the best stability,followed by RPS2.The results analyzed using geNorm and BestKeeper software are consistent.Both results showed the expression stability of the RPS2 gene is the best,followed by the 18S rRNA.Therefore,it is recommended that the RPS2+18S rRNA internal reference genes combination could be used fot the study of gene expression in different tissues of white king pigeon.

关 键 词：白羽王鸽 内参基因 表达稳定性 

分 类 号：S836.2[农业科学—畜牧学]