用于文库筛选的植物瞬时表达载体的构建

Construction of A Plant Transient Expression Vector for Library Screening

利用绿色荧光蛋白基因可以指示目的基因表达,因此本实验构建了含有稀有酶切位点的植物瞬时表达载体,并且目的基因和绿色荧光蛋白基因有各自独立的表达框架。目前在大果型的离体番茄果实上尚未有进行EGFP的瞬时表达研究,本实验利用农杆菌介导的瞬时表达技术在烟草叶片和大果型离体番茄果实上瞬时表达了绿色荧光蛋白,验证了载体进行瞬时表达的可行性。

The study of genomics depends on quick and high-flux screening of functional genes, and the rapid development of plant transient expression technology makes it possible to express exogenous genes in plants to widen the investigation of functional genes. The use of green fluorescent protein gene can indicate the expression of target gene, therefore we constructed a plant transient expression vector containing two separate expression frames, one with rare restriction enzyme cleavage sites for cloning and expression of target gene and the other flame with enhanced green fluorescent protein gene. Meanwhile, the vector transiently expressed green fluorescent protein gene in tobacco leaves and detached tomato fruits by agrobacteriurn-mediated method, indicating the potent feasibility of this vector for transient expression.