Genetic transformation of plants offers the possibility of functional characterization of individual genes and the improvement of plant traits. Development of novel transformation vectors is essential to improve plant genetic transformation technologies for various applications. Here, we present the development of a Gateway-compatible two-component expression vector system for Agrobacterium-mediated plant transformation. The expression system contains two independent plasmid vector sets, the activator vector and the reporter vector, based on the concept of the GAL4/UAS trans-activation system. The activator vector expresses a modified GAL4 protein (GAL4-VP16) under the control of specific promoter. The GAL4-VP16 protein targets the UAS in the reporter vector and subsequently activates reporter gene expression. Both the activator and reporter vectors contain the Gateway recombination cassette, which can be rapidly and efficiently replaced by any specific promoter and reporter gene of interest, to facilitate gene cloning procedures. The efficiency of the activator-reporter expression system has been assessed using agroinfiltration mediated transient expression assay in Nicotiana benthamiana and stable transgenic expression in Arabidopsis thaliana. The reporter genes were highly expressed with precise tissue-specific and subcellular localization. This Gateway-compatible two-component expression vector system will be a useful tool for advancing plant gene engineering.

中文翻译：

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开发适用于植物的网关兼容的双组分表达载体系统。


植物的遗传转化为个体基因的功能表征和植物性状的改良提供了可能性。开发新型转化载体对于改进植物遗传转化技术的各种应用至关重要。在这里，我们展示了用于农杆菌介导的植物转化的网关兼容的双组分表达载体系统的开发。基于GAL4/UAS反式激活系统的概念，表达系统包含两个独立的质粒载体组，激活载体和报告载体。激活载体在特定启动子的控制下表达修饰的 GAL4 蛋白 (GAL4-VP16)。 GAL4-VP16 蛋白靶向报告载体中的 UAS，随后激活报告基因表达。激活子和报告子载体均含有 Gateway 重组盒，可以快速有效地被任何特定的感兴趣的启动子和报告子基因替换，以促进基因克隆程序。使用农杆菌渗透介导的烟草瞬时表达测定和拟南芥中的稳定转基因表达评估了激活子-报告基因表达系统的效率。报告基因高度表达，具有精确的组织特异性和亚细胞定位。这种与网关兼容的双组分表达载体系统将成为推进植物基因工程的有用工具。