东北林蛙GSK3β基因荧光定量PCR引物筛选体系的建立  

Selection of Primers and Development of a GSK3 Beta Reaction System for Rana dybowskii

作　　者：张宇[1] 巩珊珊[1] 柴龙会[1] 张晶钰[1] 肖向红[1] 

机构地区：[1]东北林业大学野生动物资源学院,哈尔滨150040

出　　处：《野生动物学报》2016年第3期242-245,共4页CHINESE JOURNAL OF WILDLIFE

基　　金：国家级实验教学示范中心建设专项;国家自然科学基金项目(30870301);黑龙江省自然科学基金重点项目(ZJN0604-02)

摘　　要：为探究低温条件下东北林蛙(Rana dybowskii)肝脏中GSK3β基因表达量变化,需利用荧光定量PCR(Fluorescence quantitative real-time PCR)对GSK3β基因表达量进行检测,而在此过程中PCR引物的筛选直接影响到检测结果数据的准确性与真实性。本实验以东北林蛙为实验对象,基于本研究组前期获得的东北林蛙转录组数据库,并参考其他物种GSK3β基因,利用Beacon Designer 7软件设计了6对引物,采用逆转录PCR和实时荧光定量PCR技术,筛选出1对GSK3β基因荧光定量PCR引物(F-TCCTACATCTGCTCTCGGTA,R-ACATCTATGCTGGAGGTATAATCA),其扩增效率为E=99.3%、R^2=0.998,可用于东北林蛙GSK3β基因实时表达量的研究。In order to further explore the condition of GSK3- beta mRNA expression volume change in Rana dybowskii＇s liver at low temperature,we need to use the realtime fluorescent quantitative PCR（RTQF- PCR） to test the amount of GSK3- beta gene expression,and the process of PCR primers screening directly affects the authenticity and accuracy of test results.Based on the transcriptome database of Rana dybowskii,and using GSK3 beta gene of other species as reference,we designed 6 pairs of primers using the software Beacon Designer 7.These primer- pairs were screened by RT- PCR and Real- time fluorescent quantitative PCR.One pair of fluorescence quantitative PCR primers（F- TCCTACATCTGCTCTCGGTA.R- ACATCTATGCTGGAGGTATAATCA）for the GSK3 beta gene was selected.The amplification efficiency was ？ = 99.3%,and R2= 0.998.The pair of primers can be used for research on GSK3 beta gene expression by Rana dybowskii.

关 键 词：GSK3Β 荧光定量PCR 引物 

分 类 号：Q959.6[生物学—动物学]