Analysis of the spatial and temporal expression pattern directed by the Populus tomentosa 4-coumarate:CoA ligase Pto4CL2 promoter in transgenic tobacco
Analysis of the spatial and temporal expression pattern directed by the Populus tomentosa 4-coumarate:CoA ligase Pto4CL2 promoter in transgenic tobacco
复制标题
转基因烟草中毛白杨4-香豆酸辅酶A连接酶Pto4CL2启动子指导的时空表达模式分析
DOI：
10.1007/s11033-012-2312-6
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发表时间：
2013-03-01
影响因子：
2.8
通讯作者：
Lu, Hai
中科院分区：
文献类型：
--
作者：
Pan, Xiang;Li, Huanhuan;Lu, Hai
4-香豆酸：CoA连接酶（4-Coumarate：CoA ligase，4CL）是苯丙素类化合物合成途径中的关键酶。从毛白杨（Populus tomentosa）卡尔（Carr）中克隆了Pto 4 CL 2启动子。并与编码β-葡萄糖醛酸酶（GUS）的报告基因融合，通过组织化学方法对Pto 4CL 2启动子指导下的复合表达模式进行了表征。对转化体进行的启动子5 '缺失和组织化学分析表明，-317至-292 nt区域支持Pto 4CL 2在表皮和花瓣中表达，而-266至-252 nt区域的缺失导致组织特异性的丧失和GUS活性的显著降低。电泳迁移率变动分析表明，Pto 4CL 2启动子中富含腺嘌呤和胞嘧啶的元件（-264 ~-255 nt）和脱落酸响应元件（-242 ~-235 nt）分别具有复杂表达谱和有效基础表达的功能。这些结果进一步阐明了II类4CL启动子在高等植物中的调控表达模式。
4-Coumarate:CoA ligase (4CL) is a key enzyme in the phenylpropanoid synthesis pathway. The Pto4CL2 promoter was cloned from Populus tomentosa Carr. and fused to the reporter gene encoding β-glucuronidase (GUS); the complex expression patterns directed by the Pto4CL2 promoter were then characterized in Nicotiana tabacum Xanthi by histochemical assays. The promoter 5'-deletion and histochemical assay conducted on transformants indicated that the -317 to -292 nt region supports Pto4CL2 expression in the epidermis and petals and the deletion of the -266 to -252 nt region resulted in the loss of tissue specificity and a dramatic reduction in GUS activity. Furthermore, electrophoretic mobility shift assays testified that an adenine and cytosine-rich element (-264 to -255 nt) and an abscisic acid-responsive element (-242 to -235 nt) in the Pto4CL2 promoter would have functions for the complex expression profiling and efficient basal expression, respectively. These results further clarify the mode of the regulatory expression of class II 4CL promoters in higher plants.