天津农业科学2024，Vol.30Issue(7)：54-59,66,7.DOI:10.3969/j.issn.1006-6500.2024.07.009
黑线姬鼠睾丸实时荧光定量PCR内参基因的筛选及验证
Screeningand Verification of Reference Genes for Real-time Quantitative PCR Analysis in Apodemus a-grarius Testicles
摘要
Abstract
In order to screen out the optimal internal reference genes of Apodemus agrarius testicles in different seasons,this study was based on the transcriptome data of Apodemus agrarius testicles,twelve candidate reference genes were initially selected,namely AL-DOA,GLUL,RABAC1,TEGT,EID3,SYF2,MRPL20,GAPDH,HPRT,18s,B2M and NWD1.The testicles of different seasons(spring,summer and autumn)were extracted from RNA and reverse transcribed into cDNA.The expression levels of the above candi-date reference genes were then measured by Real-time Quantitative PCR.The Cyclethreshold was analyzed by the analysis software GeNorm,NormFinder,BestKeeper and RefFinder.The results showed that the expression stability of 12 candidate reference genes in the testis of Apodemus agrus in different seasons was ranked from high to low as STF2＞B2M＞GAPDH＞MRPL20＞18s＞HPRT＞ALDOA＞EID3＞RABAC1＞TEGT＞NWD1＞GLUL,among which SYF2 was the most stable reference gene,GLUL was the most unstable gene.Among them,SYF2 was the most stable reference gene,GLUL was the least unstable gene.This study provides a basis for functional gene development and gene expression research of Apodemus agrarius,and provides a reference for subsequent ge-nomics research of Apodemus agrarius.关键词
黑线姬鼠/内参基因/表达稳定性/不同季节/qRT-PCRKey words
Apodemus agrarius/reference gene/expression stability/different seasons/qRT-PCR分类
生物科学引用本文复制引用
金志民,李强,王迎新,李璐,任占辰,贾修歧..黑线姬鼠睾丸实时荧光定量PCR内参基因的筛选及验证[J].天津农业科学,2024,30(7):54-59,66,7.基金项目
国家重点研发计划子课题(2023YFF0724603-4) （2023YFF0724603-4）
黑龙江省2023年度省属高校基本科研业务费科研项目(1453QN029) （1453QN029）
牡丹江师范学院青年学术骨干项目(MNUQN202305) （MNUQN202305）
生物学学科卓越研究生培养模式改革创新项目(MSYZYXM2022004) （MSYZYXM2022004）
