水稻OsWRKY17基因定位表达载体的构建

摘要

[目的]研究水稻OsWRKY17基因的生理生化特性,确定OsWRKY17蛋白在植物中的定位.[方法]根据GenBank数据库中OsWRKY17全序列设计引物,进行Os WRKY17的RT-PCR扩增,克隆了该基因,将该片段与带绿色荧光蛋白(GFP)基因的质粒载体pBinG-FP重组,将构建正确的表达载体pBinGFP-OsWRKY17通过农杆菌介导的花蕾浸泡法转化到拟南芥中.[结果]经菌落PCR与酶切鉴定表明成功构建了OsWRKY17基因与GFP融合的植物表达载体pBinGFP-OsWRKY17,并成功将OsWRKY17基因整合到拟南芥的基因组中,获得了抗性植株.[结论]OsWRKY17基因表达载体的构建为研究该基因的生理生化特性奠定了基础.%[Objective] To study the physiological biochemical characteristic of OsWRKY17 in rice and identify the subcellular location of Os-WRKY17. [Method] The primer of the OsWRKYYl gene was designed according to the OsWRKY17 full length sequence in Genbank and cloned by RT - PCR. The cloned fragment was then recombined with the green fluorescent protein gene of plasmid vector pBinGFP. The recombinant plasmid pBinGFP - 0sWRKY17 was transformed into Arabidopsis through Agrobacterium tumefaciens strain GV3101. [Result] Colony PCR and digestion identification proved that the plant expression vector pBinGFP - OsWRKY17 was successfully constructed by the fusion of Os-WRKY1 and GFP,and the expression vector was successfully transformed into the genome of Arabidopsis,obtaining resistant plant. [Conclusion] Construction of OsWRKYM expression vector established the foundation for study the physiological biochemical characteristics of Os-WRKY17.