Construction vascular-specific expression bi-directional promoters in plants
Construction vascular-specific expression bi-directional promoters in plants
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构建植物维管特异性表达双向启动子
DOI：
10.1016/j.jbiotec.2009.03.009
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发表时间：
2009-05-20
影响因子：
4.1
通讯作者：
Lu, Hai
中科院分区：
文献类型：
--
作者：
Lv, Xiaomeng;Song, Xiaodan;Lu, Hai
已广泛用于植物基础研究和生物技术应用的启动子通常是单向的。在这里，我们描述了一种策略，双向化的血管特异性表达grp1.8启动子（这里命名为GRPp）和4CL 1启动子（这里命名为4CL 1 p），使一个启动子可以指导两个基因的血管特异性表达，一个在启动子的两端。将最小启动子（35 Smini或GRP mini）与特异表达启动子（GRPp或4CL 1 p）的5'端融合形成双向启动子（35 Smini-GRPp、35 Smini-4CL 1 p或GRPnini-GRPp），可在所有独立的转基因烟草株系中指导葡萄糖醛酸苷酶（gus）和绿色荧光蛋白（gfp）基因的表达。通过GUS组织化学染色和荧光显微镜下观察转基因植株中GFP的表达，分析了gusA和gfp基因在转基因植株中的稳定表达。荧光定量PCR检测到GFP和GUS基因在转基因烟草中的转录水平显著提高。它们的维管特异性双向启动子可用于同时在转基因植物中特异表达多个功能基因。(C)2009 Elsevier BY. All rights reserved.
Promoters that have been widely used for both basic research and biotechnological application in plants are generally unidirectional. Here we describe a strategy to bi-directionalize the vascular-specific expression grp1.8 promoter (here named GRPp) and 4CL1 promoter (here named 4CL1p) so that one promoter can direct the vascular-specific expression of two genes, one on each end of the promoter. The minimal promoter (35Smini or GRP mini), when fused at the 5' end of the specific expression promoter (GRPp or 4CL1p) to form bi-directional promoter (35Smini-GRPp, 35Smini-4CL1p or GRPnnini-GRPp), was able to direct expression of the glucuronidase (gus) and green fluorescent protein (gfp) gene in all independent transgenic tobacco lines. Stable expression of gusA and gfp genes in transgenic plants was analyzed by histochemical staining for GUS and fluorescence microscopic observation under UV for GFP in transgenic plants. The remarkable transcript levels of GFP and GUS were detected by real-time PCR in independent transgenic tobacco lines. Their vascular-specific bi-directional promoters should be used to vascular-specific expression several functional genes in transgenic plants simultaneously. (C) 2009 Elsevier BY. All rights reserved.