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一种结合了花椰菜花叶病毒35S启动子和甘露氨酸合成酶基因启动子元件的嵌合植物启动子的新颖且有用的特性。

Novel and useful properties of a chimeric plant promoter combining CaMV 35S and MAS elements.

作者信息

Comai L, Moran P, Maslyar D

机构信息

Calgene Inc, Davis, CA 95616.

出版信息

Plant Mol Biol. 1990 Sep;15(3):373-81. doi: 10.1007/BF00019155.

Abstract

The CaMV 35S and Ti plasmid mannopine synthetase (mas) promoters are commonly used by plant genetic engineers. To combine their useful properties, we constructed hybrid promoters incorporating elements from both. These promoters were spliced to the beta-glucuronidase reporter gene and introduced into tobacco and tomato plants by Agrobacterium cocultivation. T1 and T2 transgenic plant populations transformed with different constructs were assayed for the marker enzyme. Comparisons were made based on the range of expression levels found for each promoter construct. We found that a hybrid promoter incorporating the mas region from +65 to -301 and the 35S enhancer region from -90 to -941 had new and interesting properties. This promoter, called Mac, expressed gus at a level three to five times that expressed by a double 35S promoter in the leaves, and 10 to 15 times in hypocotyls and roots. The Mac promoter, however, showed only marginal wound inducibility. Five- to seven-fold wound induction required the presence of the region from -301 to -613 of mas. Reiteration of the 35S enhancer region, from -90 to -430, behind the 35S TATA box region or the mas +65 to -301 region had a smaller effect on expression, ranging from equal to twice the level of the single enhancer control.

摘要

花椰菜花叶病毒35S启动子(CaMV 35S)和Ti质粒甘露碱合成酶(mas)启动子是植物基因工程中常用的启动子。为了结合它们的有用特性,我们构建了包含两者元件的杂交启动子。这些启动子与β-葡萄糖醛酸酶报告基因拼接,并通过农杆菌共培养导入烟草和番茄植株。对用不同构建体转化的T1和T2转基因植株群体进行了标记酶检测。根据每个启动子构建体的表达水平范围进行了比较。我们发现,一个包含从+65到-301的mas区域和从-90到-941的35S增强子区域的杂交启动子具有新的有趣特性。这个启动子称为Mac,在叶片中驱动gus表达的水平是双35S启动子的三到五倍,在下胚轴和根中是其10到15倍。然而,Mac启动子仅表现出微弱的创伤诱导性。五到七倍的创伤诱导需要mas的-301到-613区域的存在。在35S TATA框区域或mas的+65到-301区域之后重复35S增强子区域(从-90到-430)对表达的影响较小,范围从等于单增强子对照水平的两倍。

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