Genetically encoded fluorescent proteins are often used to label proteins and study protein function and localization in vivo. Traditional cloning methods mediated by restriction digestion and ligation are time-consuming and sometimes difficult due to the lack of suitable restriction sites. Invitrogen developed the Gateway cloning system based on the site-specific DNA recombination, which allows for digestion-free cloning. Most gateway destination vectors available for use in plants employ either the 35S or ubiquitin promoters, which confer high-level, ubiquitous expression. There are far fewer options for moderate, cell-type specific expression. Here we report on the construction of a Gateway-compatible cloning system (SWU vectors) to rapidly tag various proteins and express them in a cell-type specific manner in plants. We tested the SWU vectors using the HISTONE (H2B) coding sequence in stable transgenic plants. The SWU vectors are a valuable tool for low cost, high efficiency functional analysis of proteins of interest in specific cell types in the Arabidopsis root.

中文翻译：

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用于以细胞类型特异性方式对蛋白质进行功能分析的网关兼容载体。

基因编码的荧光蛋白通常用于标记蛋白质和研究蛋白质功能和体内定位。由限制性消化和连接介导的传统克隆方法耗时且有时由于缺乏合适的限制性位点而困难。Invitrogen 开发了基于位点特异性 DNA 重组的 Gateway 克隆系统，可实现免消化克隆。大多数可用于植物的网关目标载体采用 35S 或泛素启动子，它们可提供高水平、无处不在的表达。适度的细胞类型特异性表达的选择要少得多。在这里，我们报告了网关兼容克隆系统（SWU 载体）的构建，以快速标记各种蛋白质并在植物中以细胞类型特异性方式表达它们。我们在稳定的转基因植物中使用组蛋白 (H2B) 编码序列测试了 SWU 载体。SWU 载体是一种有价值的工具，可用于对拟南芥根中特定细胞类型中感兴趣的蛋白质进行低成本、高效率的功能分析。