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首页> 外文期刊>Journal of Biotechnology >Construction vascular-specific expression bi-directional promoters in plants.
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Construction vascular-specific expression bi-directional promoters in plants.

机译:在植物中构建血管特异性表达双向启动子。

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摘要

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 GRPmini-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.
机译:已经广泛用于植物的基础研究和生物技术应用的启动子通常是单向的。在这里,我们描述了一种双向化血管特异​​性表达 grp1.8 启动子(此处称为 GRPp )和 4CL1 启动子(此处称为战略)的策略 4CL1p ),以便一个启动子可以指导两个基因的血管特异性表达,每个基因在启动子的每一端。最小启动子( 35Smini 或 GRP mini )在特定表达启动子( GRPp 或 4CL1p)的5'端融合时)形成双向启动子( 35Smini-GRPp , 35Smini-4CL1p 或 GRPmini-GRPp ),葡萄糖醛酸酶( gus )和绿色荧光蛋白( gfp )基因在所有独立的转基因烟草品系中的表达通过GUS的组织化学染色和UV下的荧光显微镜观察转基因植物中的GFP,分析了转基因植物中 gusA 和 gfp 基因的稳定表达。通过实时PCR在独立的转基因烟草品系中检测到了 GFP 和 GUS 的显着转录本水平。它们的血管特异性双向启动子应用于在转基因植物中同时特异性表达几种功能基因。

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