白羽绿头鸭骨骼肌发育内参基因的鉴定和筛选  

Identification and screening of reference genes for skeletal muscle development in white-feathered mallard

作　　者：赵明明 刘博 吴琼 赵家平[1] 涂剑锋[1] ZHAO Mingming;LIU Bo;WU Qiong;ZHAO jiaping;TU Janfeng(Institute of Special Animal and Plant Science,Chinese Academy of Agricultural Sciences,Changchun 130117,China;College of Life Sciences,Longyan University,Longyan 364012,China)

机构地区：[1]中国农业科学院特产研究所,长春130117 [2]龙岩学院生命科学学院,福建龙岩364012

出　　处：《黑龙江畜牧兽医》2021年第23期122-129,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：吉林省科技发展计划项目(2018020132NY)。

摘　　要：为选取适宜进行白羽绿头鸭骨骼肌发育相关基因研究的内参基因,试验通过实时荧光定量聚合酶链式反应(RT-qPCR)对13个候选内参基因进行表达稳定性验证,利用geNorm、NormFinder软件评估候选内参基因稳定性,利用BestKeeper软件对geNorm和NormFinder软件评估结果中稳定性排前10位的候选内参基因进行稳定性分析,用RefFinder软件对内参基因稳定性进行综合评价,并通过geNorm软件确定最适内参基因数量。结果表明:geNorm软件评估结果显示SEC61G与NDUFB3组合内参基因稳定性最高,HPRT1基因稳定性最低,最适内参基因数为2个;NormFinder软件评估结果显示NDUFB3稳定性最高,HPRT1基因稳定性最低;BestKeeper软件评估结果显示SAT1稳定性最高,GAPDH基因稳定性最低;Refinder软件评估结果显示候选内参基因稳定性从高到低依次为NDUFB3>SEC61G>SOD1>SAT1>TOP2B>ATP5MF>HMBS>HINT1>GAPDH>YWHAZ>RPS2>ACTB>HPRT1,HPRT1的表达最不稳定,NDUFB3基因表达最稳定。说明在白羽绿头鸭骨骼肌发育中,最佳内参基因为NDUFB3,最佳内参基因组合为NDUFB3和SEC61G,2个内参基因最佳。In order to select suitable reference genes for the study of genes related to skeletal muscle development in white-feathered mallard,the expression stability of 13 candidate reference genes was verified by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).GeNorm and NormFinder software were used to evaluate the stability of candidate reference genes.BestKeeper software was used to analyze the stability of the top 10 candidate internal reference genes in the evaluation results of geNorm and NormFinder software.RefFinder software was used to comprehensively evaluate the stability of internal reference genes.GeNorm software was used to determined the optimal number of reference genes.The results showed as follows:the evaluation results of geNorm software showed that the combination of SEC61 G and NDUFB3 had the highest stability.The stability of HPRT1 was the lowest,and the optimum number of reference genes was 2.NormFinder software showed the highest stability of NDUFB3 and the lowest stability of HPRT1.BestKeeper softwere showed that SAT1 had the highest stability and GAPDH had the lowest stability.RefFinder softwere showed that the stability of candidate internal reference genes from high to low was:NDUFB3>SEC61 G>SOD1>SAT1>TOP2 B>ATP5 MF>HMBS>HINT1>GAPDH>YWHAZ>RPS2>ACTB>HPRT1.Among them,the expression of HPRT1 was the most unstable,and the reference gene with the most stable expression was NDUFB3.These results indicated that the best reference gene was NDUFB3,the best combination of reference genes was NDUFB3 and SEC61 G,and two reference genes were the best.

关 键 词：白羽绿头鸭 内参基因 骨骼肌发育 稳定性 筛选 

分 类 号：S834.89[农业科学—畜牧学] Q344.13[农业科学—畜牧兽医]