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[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OsWRKY17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] The research aimed to study the physiological and biochemical characteristics of OsWRKY17 in rice and determine the localization of OsWRKY17 in plants. [Method] According to the complete sequence of OsWRKY17 in GenBank, primers were designed and OsWRKY17 was amplified by RT-PCR. OsWRKY17 gene was cloned and recombined with plasmid pBinGFP with green fluorescent protein (GFP) gene to construct the correct expression Vector pBinGFP-OsWRKY17 was transformed into Arabidopsis by Agrobacterium-mediated bud dipping. [Result] The results of colony PCR and restriction enzyme digestion indicated that the plant expression vector pBin-GFP / OsWRKY17 fused with GFP was successfully constructed and the OsWRKY17 gene was successfully integrated into the genome of Arabidopsis. The resistant plants . [Conclusion] The construction of OsWRKY17 gene expression vector laid the foundation for studying the physiological and biochemical characteristics of this gene.