东北林蛙GSK3β基因荧光定量PCR引物筛选体系的建立

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-ACATCTA...

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Bibliographic Details
Published in野生动物学报 Vol. 37; no. 3; pp. 242 - 245
Main Author 张宇 巩珊珊 柴龙会 张晶钰 肖向红
Format Journal Article
LanguageChinese
Published Editorial Department of Chinese Journal of Wildlife 2016
Subjects
GSK3β
引物
荧光定量PCR
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ISSN2310-1490

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Summary: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。
Bibliography:GSK3-beta; Fluorescence quantitative real-time PCR; Primers
Zhang Yu, Gong Shanshan ,Chai Longhui , Zhang Jingyu, Xiao Xianghong （College of Wildlife Resources, Northeast Forestry University, Haerbin, 150040, China）
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= 0.9
ISSN:2310-1490