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，进行仉WRKY]7的RT—PCR扩增，克隆了OsWRKY17基因，将该片段与带绿色荧光蛋白（GFP）基因的质粒载体pBinGFP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Os—WRKY17基因与GFP融合的植物表达载体pBin—GFP／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 Os- WRKY17 in rice and identify the subcellular location of OsWRKY17. [Method] The primer of the OsWRKY17 gene was designed according to the full-length sequence of OsWRKY17 in Genbank and was 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 diges- tion 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, there by ob- taining a resistant plant. [Conclusion] The construction of OsWRKY17 expression vector established the foundation for study on the physiological the biochemical char- acteristics of QsWRKY17.