《植物生理学报》 2015, 51(4): 495-500

三七鲨烯环氧酶基因(SE)启动子的瞬时表达分析

卢荣江, 刘迪秋, 葛锋*, 陈朝银

昆明理工大学生命科学与技术学院, 昆明650500

通信作者：葛锋；E-mail: gefeng@tsinghua.org.cn；Tel: 0871-65920621

摘　要：

采用PCR方法扩增鲨烯环氧酶(SE)基因启动子序列, 研究SE基因在三七生物合成途径中的表达调控规律。为了确认启动子表达的最佳启动区域, 构建SE启动子430、922、1 323和1 872 bp的5'端缺失片段, 并将这些缺失片段分别与葡萄糖苷酸酶(GUS)基因融合, 构建植物表达载体。采用农杆菌介导法转化烟草, 在烟草叶片中进行瞬时表达分析。组织化学染色和GUS荧光定量分析结果显示, 4个不同长度片段的启动子均有启动活性, 且随着缺失长度增加启动活性呈增加趋势。

关键词：三七; 鲨烯环氧酶; 启动子; 瞬时表达

收稿：2014-12-11   修定：2015-03-06

资助：国家自然科学基金项目(31260070)。

Transient Expression Analysis of Squalene Epoxidase Gene (SE) Promoter from Panax notoginseng

LU Rong-Jiang, LIU Di-Qiu, GE Feng*, CHEN Chao-Yin

Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China

Corresponding author: GE Feng; E-mail: gefeng@tsinghua.org.cn; Tel: 0871-65920621

Abstract:

In order to study the expression and regulation of squalene epoxidase (SE) gene in the biosynthesis pathway in Panax notoginseng, the SE promoter was cloned by PCR method. To determine the optimal promoter sequence for gene expression, SE promoter was deleted from its 5' end to form promoter fragments with 430, 922, 1 323 and 1 872 bp. Such fragments were fused to a β-glucuronidase (GUS) gene. The fused genes were transformed into Nicotiana tabacum using Agrobacterium tumefaciens-mediated method for transient expression. Histochemical staining and GUS quantitative fluorometric assays demonstrated that the four different length fragments of the promoter had promoter activities and the activities were increased with the deleted length of promoters.

Key words: Panax notoginseng; squalene epoxidase; promoter; transient expression