利用双荧光报告基因可视化鉴定胚特异启动子的功能

Using Dual Fluorescent Reporter Genes to Visually Identify the Function of Embryo-Specific Promoters

胚特异启动子对于外源基因在种子中特异表达、改良水稻品质有重要理论和实际意义。为高效鉴定水稻胚特异启动子功能和表达情况,本实验筛选了三个胚特异启动子OsESP1、pEMB1和Ole18,分别连接到含红色荧光蛋白(RFP)报告基因的载体pLJ02上构建双荧光表达载体。利用基因枪介导的瞬时表达结合农杆菌介导的稳定表达,通过荧光观察,鉴定三个胚特异启动子的特异表达模式。结果表明:GFP/RFP双荧光载体是一个可以用于可视化筛选组织特异启动子的高效表达载体;相较于成熟胚,幼胚更适宜用于胚特异表达启动子的瞬时表达验证,为研究组织特异性启动子驱动外源基因在水稻中表达及水稻品种遗传改良拓展新思路。

Embryo-specific promoters have important theoretical and practical significance for the specific expression of foreign genes in seeds and the improvement of rice quality.In order to efficiently identify the function and expression of rice embryo-specific promoters,three embryo-specific promoters OsESP1,pEMB1 and Ole18 were screened in this experiment,and they were connected to the vector pLJ02 containing the red fluorescent protein(RFP)reporter gene to construct a dual fluorescent expression vector.Using gene gun-mediated transient expression combined with Agrobacterium-mediated stable expression,through fluorescence observation,the specific expression patterns of the three embryo-specific promoters were identified.The results showed that the GFP/RFP dual fluorescent vector is a highly efficient expression vector that can be used to visually screen tissue-specific promoters;compared to mature embryos,immature embryos are more suitable for the transient expression verification of embryo-specific expression promoters.To develop new ideas for the study of tissue-specific promoters driving the expression of foreign genes in rice and the genetic improvement of rice varieties.