岗梅实时荧光定量PCR内参基因的筛选

Selection of Reference Gene for Real-time PCR Study in Ilex asprella

为筛选适用于岗梅实时荧光定量PCR的内参基因。应用实时荧光定量PCR技术,分析18S r RNA(18S)、Actin(ACT)、α-tubulin(TUA)、β-tubulin(TUB)、Cyclophili(CYP)、Elongation factor 1-α(EF-1α)和Polyubiquitin(UBQ)共7个看家基因在岗梅的8个不同组织部位中的表达情况,并利用Ge Norm、Norm Finder和Best Keeper共3个软件对每个看家基因的表达稳定性进行评价。3个软件的分析结果均表明:18S、ACT和TUA看家基因的稳定性和相关性均较好,可在这3个看家基因中选择1个或以18S-ACT、18S-TUA组合作为岗梅基因研究的内参基因。

This study aimed to select reference gene （s）for real time quantitative PCR （qRT-PCR） research in I. asprella. The expression levels of seven housekeeping genes （HKGs）, including 18S rRNA （18S）, Actin（ACT）, α- Tubulin （ TUA ）, β-Tubulin （TUB）, Cyclophili （ CYP）, Elongation factor 1-α （ EF 1α ） and Polyubiquitin （UBQ）, in eight different tissues of I. asprella, were analyzed by qRT-PCR. Furthermore, their expression stabilities were evaluated by three software, including GeNorm, NormFinder and BestKeeper. Analysis data showed that 18S, ACT and TUA expressed stably and were related to each other in different tissues of I. asprella. 18S, A CT and TUA, combination of 18S-ACT or 18S-TUA, could be considered as candidate reference gene（s） for qRT-PCR study of 1. asprella.