作 者：
赵晓冰;潘磊;柳威;林懋怡;李宏庆;刘忠
关键词：
华细辛;内参基因;geNorm;Norm Finder;Best Keeper;Delta CT
摘 要：
选择合适的内参基因是准确分析目标基因表达水平变化的重要条件。选取SAND-1、ACT、18Sr RNA、CYP2、GAPDH、TUB 6个常用的内参基因作为候选内参基因,利用实时荧光定量PCR技术,通过ge Norm、Norm Finder、Best Keeper、Delta CT等软件分析和评价6个候选内参基因在华细辛营养生长期、花期和花后期等不同生长阶段的根、根茎、叶片、叶柄等不同组织中的表达稳定性,筛选适宜华细辛不同生长阶段不同组织基因表达水平分析的内参基因。结果表明,18S r RNA在华细辛所有样品中表达最为稳定,是进行华细辛基因表达水平分析的适宜的内参基因。研究结果可为华细辛重要活性成分生物合成途径相关基因的功能分析,以及基因差异表达的研究提供有效的校正工具,确保基因表达分析结果的准确性与可靠性。
译 名：
Screening of Reference Genes for Quantitative Real-time PCR Analysis in Asarum sieboldii
作 者：
ZHAO Xiao-bing;PAN Lei;LIU Wei;LIN Mao-yi;LI Hong-qing;LIU Zhong;School of Life Science,East China Normal University;School of Pharmacy,Shanghai Jiao Tong University;
单 位：
School of Life Science,East China Normal University%School of Pharmacy,Shanghai Jiao Tong University
关键词：
Asarum sieboldii;;reference gene;;GeNorm;;Norm Finder;;Best Keeper;;Delta CT
摘 要：
Selecting appropriate reference gene is key step for the accurate analysis of the expression variations of target genes. Usingseveral common-used reference genes of SAND-1,ACT,18 S r RNA,CYP2,GAPDH and TUB as candidate genes and quantitative realtime PCR analysis of gene expression,the expression stabilities of these 6 candidate genes in various samples from roots,rhizomes,leaves,petioles and flowers in different growth stages of nutrition growth,flowering and post-flowering were assessed,with a set of approaches suchas ge Norm,Norm Finder,Best Keeper and Delta CT,representatively. It is for screening the appropriate reference genes in analysing geneexpressions of Asarum sieboldii Miq. in varied tissues at different growth stage. The results demonstrated that 18 S r RNA was the optimal objectto act as a proper reference gene in this species because it displayed the most stable expression in almost all samples. This study will provide afeasible and valid calibrate not only for gene function characterization concerning chemical compound biosynthesis but also for gene differentialexpression analysis in A. sieboldii,ensuring the accuracy and reliability of results.
