水稻OsWRKY17基因定位表达载体的构建

Construction of OsWRKY17 Specific Expression Vector in Plant

[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Os-WRKY17全序列设计引物,进行OsWRKY17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 OsWRKY17.[Method] The primer of the OsWRKY17 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-OsWRKY17 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 OsWRKY17 and GFP,and the expression vector was successfully transformed into the genome of Arabidopsis,obtaining resistant plant.[Conclusion] Construction of OsWRKY17 expression vector established the foundation for study the physiological biochemical characteristics of OsWRKY17.