《植物生理学报》 2016, 52(12): 1927-1934

拟南芥维管组织特异性启动子的人工构建及活性检测

刘石娟^*, 王雪

曲阜师范大学生命科学学院, 山东曲阜273165

通信作者：刘石娟；E-mail: sjliu@mail.qfnu.edu.cn

摘　要：

利用组织特异性启动子驱动外源基因在维管中特异表达, 可以更有效地发挥目的基因防治病虫害的作用。在本研究中, 将维管组织特异性元件序列的串联拷贝与拟南芥蛋白酪氨酸磷酸酶1基因的缺失启动子融合, 然后通过GUS报告基因检测串联拷贝及融合启动子活性。GUS组织化学染色结果表明组织特异性元件的串联拷贝能够显著提高启动子活性, 融合启动子在转基因拟南芥维管组织中特异性高效表达。GUS酶活分析和QRT-PCR实验进一步证实该融合启动子的高表达活性, 从而为抗维管束病虫害植物基因工程提供优良的调控元件。

关键词：启动子; 顺式作用元件; 组织特异性; 维管组织

收稿：2016-08-15   修定：2016-11-13

资助：国家自然科学基金(31670270和31200196)和山东省自然科学基金(ZR2011CQ016)。

Construction and functional analysis of a synthetic vascular-specific promoter in Arabidopsis

LIU Shi-Juan^*, WANG Xue

College of Life Sciences, Qufu Normal University, Qufu, Shandong 273165, China

Corresponding author: LIU Shi-Juan; E-mail: sjliu@mail.qfnu.edu.cn

Abstract:

Precise control of transgene expression from vascular-specific promoters is preferred for the production of engineered plants with pests and disease resistance. In this study, a synthetic plant promoter was created by combining two copies of the vascular-specific elements with the truncated promoter of the Arabidopsis protein tyrosine phosphatase 1 gene. The activity of this synthetic promoter was estimated by using promoter-GUS transgene approach. GUS histochemical staining showed that direct repeats of vascular-specific elements caused a large increase in promoter activity and the synthetic promoter directed vascular-specific expression in the transgenic Arabidopsis thaliana plants. GUS fluorometric analysis and QRT-PCR assay further confirmed the high activity of the fusion promoter, which provides novel promoter elements for engineering pests and disease resistance in crops.

Key words: promoter; cis acting element; tissue-specific; vascular tissue