适于植物转基因体系优化的绿色荧光蛋白表达载体构建及其瞬时表达验证

在建立和优化植物的转基因体系时，通常期望转化载体同时具有适用植株范围广、尺寸较小、具有多重选择标记或报告基因等优点。为此，通过Gateway技术，构建了含有绿色荧光蛋白基因GFP的植物表达载体pGWB14-GFP，并经PCR鉴定和测序验证了序列的准确性。利用基因枪转化法，将该载体转入洋葱表皮和小麦叶片中，通过荧光显微镜检测，发现绿色荧光蛋白在洋葱表皮细胞和小麦叶片表皮细胞中均能瞬时表达，表明pGWB14-GFP在双子叶和单子叶植物中都能正常发挥功能。本载体在双子叶和单子叶植物植物中都可使用，大小低于17 kb，含有GFP报告基因，具有两套卡那霉素和潮霉素选择标记基因，用于在细菌和植物中筛选，为植物转基因体系的构建和优化提供了功能强大的转化载体。

Construction and transiently-expressed validation of the GFP expression vector for optimization of plant transformation system

Abstract: Expression vectors are important materials in establishment of plant transformation system. In this study, we constructed a GFP expression vector pGWB14-GFP through Gateway cloning technology. Then we transiently transformed this vector into Onion peels and wheat leaves by gene gun method and GFP expression was detected by epi-fluorescence microscope. The results showed that GFP can function as usual in cells of both onion and wheat, suggesting this vector can be used in the transformations of both dicot and monocot plants. This vector cotains GFP expression cassette and selection markers for kanamycin, and can be used in both dicot and monocot plants. With these advantages, this vector would become a powerful tool for the establishment of plant transformation system.