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茶愈伤组织实时定量PCR分析中内参基因的选取
史成颖,李正国,徐乾,宛晓春
0
(安徽农业大学农业部、教育部茶叶生物化学和生物技术重点实验室,合肥 230036)
摘要:
为了选取合适的内参基因来分析培养基中前体物诱导及对照的茶愈伤组织中茶氨酸代谢相关基因的差异表达,利用GenBank上登录的茶树基因序列以及通过构建文库测序所得的基因(具有完整的阅读框)共7个持家基因设计引物。在分析这些引物的扩增效率和特异性后,测定了它们在茶愈伤组织生长过程中(对照和愈伤培养基中添加含氮外源物的情况下)的表达水平。利用geNorm 和NormFinder软件分析了这些持家基因的稳定性,确定在该条件下合适的内参基因为β-actinGAPDH
关键词:  实时定量PCR  内参基因  基因表达  茶树
DOI:10.13610/j.cnki.1672-352x.20141029.021
投稿时间:2014-10-13
基金项目:国家自然科学基金(31170283), 教育部博士点基金(20113418130001), 安徽省自然科学基金(KJ2010A106)和安徽农业大学稳定人才项目(wd2011-16)共同资助。
Reference gene selection for real-time PCR normalization in tea callus
SHI Chengying,LI Zhengguo,XU Qian,WAN Xiaochun
(Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture and Ministry of Education, Anhui Agricultural University, Hefei 230036)
Abstract:
In order to obtain suitable reference genes for analysis of the related gene expression in calli of the control and treatment groups, 7 housekeeping genes from the GenBank and ESTs of cDNA library of Camellia sinensis young roots(with complete open reading frame)were chosen for primer design. After analysis the amplification efficiency and primers specificity, the expression levels of the 7 housekeeping genes in tea calli of the control and treatment groups at different growth stages were determined, and their stabilities of expressions were analyzed using geNorm and NormFinder softwares. As a result, β-actin and GAPDH are suitable reference genes under this condition.
Key words:  real -time PCR  reference gene  gene expression  Camellia sinensis

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