铁皮石斛实时定量PCR内参基因的筛选

被引：36

作者：

张岗 ^{[1
,2
]}

赵明明 ^{[1
]}

张大为 ^{[1
]}

郭顺星 ^{[1
]}

机构：

[1] 中国医学科学院-北京协和医学院药用植物研究所

[2] 陕西中医学院药学院,陕西省中药基础与新药研究重点实验室

来源：

中国药学杂志 | 2013年 / 48卷 / 19期

关键词：

铁皮石斛; 内参基因; 定量PCR; 基因表达;

D O I：

暂无

中图分类号：

S567.239 [];

学科分类号：

摘要：

目的筛选铁皮石斛(Dendrobium officinale)实时定量PCR(real time quantitative PCR,qPCR)最佳内参基因。方法采用同源克隆法、RT-PCR分离候选内参基因;qPCR分析基因的引物扩增效率及相对表达量;geNorm分析内参基因的表达稳定性;qPCR检测法呢基焦磷酸合酶(farnesyl pyrophosphate synthase,FPS)基因的表达模式。结果分离到铁皮石斛ACT-1、ACT-2、EF-1α、GAPDH、β-TUB、18S rRNA、26S rRNA等7个候选内参基因片段,引物扩增效率分别为2.12、1.98、2.03、2.08、1.97、2.18和2.05;geNorm分析表达稳定性大小为EF-1α/18S rRNA﹥ACT-1﹥ACT-2﹥GAPDH﹥β-TUB﹥26S rRNA;以EF-1α/18S rRNA为标准分析FPS基因的相对表达量大小为种子>根>原球茎>茎>叶。结论确定珍稀濒危兰科药用铁皮石斛qPCR分析的最佳内参基因,为基因表达研究提供依据。

引用

页码：1664 / 1668

页数：5

共 10 条

[1] 铁皮石斛钙依赖蛋白激酶基因的分子克隆及特征分析
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张大为
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