摘要
为筛选出青鲫不同组织中的最适内参基因,本研究利用实时荧光定量PCR(quantitative real-time polymerase chain reaction,qRT-PCR)技术检测β肌动蛋白(β-actin)、甘油醛-3-磷酸脱氢酶(GAPDH)、18S核糖体RNA(18S r RNA)和核糖体蛋白L13(RPL13)4个候选内参基因在青鲫脑、鳃、性腺、肾脏、肝脏、肌肉和脾脏7个组织中的表达情况,并利用ge Norm、NormFinder和BestKeeper等程序分析候选基因的表达稳定性。结果显示,4个内参基因在各组织的Ct值高低顺序依次为:β-actin>GAPDH>18S r RNA>RPL13;4个内参基因在不同组织的稳定性有所不同,综合评价分析显示其稳定性排序为RPL13>18S r RNA>β-actin>GAPDH,RPL13适合作为青鲫不同组织qRT-PCR分析的内参基因。本研究结果可为后续青鲫功能基因表达特征的研究提供技术支撑。
In order to select the most stable internal reference genes of Carassius auratus indigentiaus for real-time quantitative PCR,the mRNA expressions of four candidate reference genes(GAPDH、β-actin、18S rRNA and RPL13)in different tissues were evaluated.The expression stability of the four reference genes was evaluated by geNorm,NormFinder and BestKeeper.The results showed that the order of Ct values of four reference genes in each tissue wasβ-actin>GAPDH>18S rRNA>RPL13.The stability of four reference genes in different tissues was different,indicating that the stability of internal reference genes was tissue-specific.Comprehensive analysis of the three procedures showed that the expression stability of four genes was RPL13>18S rRNA>β-actin>GAPDH,and RPL13 was the most stable and most suitable for the reference gene in Carassius auratus indigentiaus tissue.The results of this study provide the basis for further exploration of the expression features of functional genes in this species.
作者
曾丹
李菁菁
周馨雨
谢成辉
唐伟伟
李林
杨品红
ZENG Dan;LI Jingjing;ZHOU Xinyu;XIE Chenghui;TANG Weiwei;LI Lin;YANG Pinhong(College of Life and Environmental Sciences,Hunan University of Arts and Science,Changde 415000,China;Hunan Provincial Collaborative Innovation Center for Efficient and Health Production of Fisheries,Hunan Provincial Key Laboratory for Health Aquaculture and Product Processing in Dongting Lake Area,Hunan Provincial Key Laboratory for Molecular Immunity Technology of Aquatic Animal Diseases,Hunan Engineering Research Center of Aquatic Organism Resources and Environmental Ecology,Zoology Key Laboratory of Hunan Higher Education,Changde Research Center for Agricultural Biomacromolecule,Changde 415000,China)
出处
《湖南文理学院学报（自然科学版）》
CAS
2024年第3期50-57,共8页
Journal of Hunan University of Arts and Science(Science and Technology)
基金
湖南文理学院博士启动项目(21BSQD12)
湖南省教育厅一般项目(22C0385)。
关键词
青鲫
内参基因
qRT-PCR
表达稳定性
Carassius auratus indigentiaus
reference gene
real-time quantitative PCR
gene expression stability
