Construction of OsWRKY17 specific expression vector in plant | 水稻OsWRKY17基因定位表达载体的构建
2012
Wang Xiaolan, Guangzhou University, Guangzhou (China), College of Life Science | Tang Xin, Xinjiang University, Urumqi (China), College of Life Science and Technology | Liu Zhongyuan, Xinjiang University, Urumqi (China), College of Life Science and Technology
Chinese. [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。
Show more [+] Less [-]English. [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.
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