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COMPARISON OF DIFFERENT CONSTITUTIVE AND INDUCIBLE PROMOTERS FOR THE OVEREXPRESSION OF TRANSGENES IN ARABIDOPSIS THALIANA

机译:拟南芥中转基因过表达的不同组成型和诱导型启动子的比较

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摘要

We compared the organ specificity and the strength of different constitutive (CaMV-35S, CaMV-35Somega, Arabidopsis ubiquitin UBQ1, and barley leaf thionin BTH6 promoter) and one inducible promoter (soybean heat-shock promoter Gmhsp17.3) in stably transformed Arabidopsis thaliana plants. For this purpose we constructed a set of plant expression vectors equipped with the different promoters. Using the uidA reporter gene we could show that the CaMV-35S promoter has the highest expression level which was enhanced two- to threefold by the addition of a translational enhancer (TMV omega element) without altering the organ specificity of the promoter. The barley leaf thionin promoter was almost inactive in the majority of lines whereas the ubiquitin promoter exhibited an intermediate strength. The heat-shock promoter was inducible up to 18-fold but absolute levels were lower than in the case of the ubiquitin promoter. Conclusive quantitative results for different organs and developmental stages were obtained by the analysis of 24 stably transformed lines per promoter construct.
机译:我们比较了稳定转化的拟南芥中不同组成成分(CaMV-35S,CaMV-35Somega,拟南芥泛素UBQ1和大麦叶硫蛋白BTH6启动子)和一个诱导型启动子(大豆热休克启动子Gmhsp17.3)的器官特异性和强度。植物。为此目的,我们构建了一组装备有不同启动子的植物表达载体。使用uidA报告基因,我们可以证明CaMV-35S启动子具有最高的表达水平,通过添加翻译增强子(TMVω元素)可以将其表达增强两倍至三倍,而不会改变启动子的器官特异性。大麦叶硫蛋白启动子在大多数品系中几乎没有活性,而遍在蛋白启动子表现出中等强度。热休克启动子可诱导高达18倍,但绝对水平低于泛素启动子。通过分析每个启动子构建体的24个稳定转化的品系,获得了不同器官和发育阶段的最终定量结果。

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