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机构地区:[1]长江大学农学院,湖北荆州434025 [2]暨南大学医药生物技术研究开发中心,广州510632
出 处:《中国农学通报》2010年第14期74-77,共4页Chinese Agricultural Science Bulletin
基 金:国家自然科学基金"玉米Pto基因的克隆及其调控网络解析"(30700433)
摘 要:Pti基因不但在植物的抗病信号传导中起着非常重要的作用,而且玉米中一个类Pti1基因——ZmPti1还参与了植物抗盐的信号传递。为研究ZmPti1在植物体内亚细胞水平上的分布情况,进而对ZmPti1基因功能提供更全面的信息,利用绿色荧光蛋白基因(GFP)为报告基因,分别构建了以35S为启动子表达ZmPti1-GFP融合蛋白和GFP游离蛋白的植物表达载体pGreen0029-35S∷ZmPti1-GFP和pGreen0029-35S∷GFP。构建好的植物表达载体首先转化感受态的农杆菌菌株GV3101,再通过花浸蘸法侵染拟南芥,结果得到62株转基因拟南芥株系。在共聚焦激光扫描显微镜下观察ZmPti1在转基因拟南芥细胞中的定位。结果表明:ZmPti1是一个可溶性蛋白,定位在细胞质内。Pti1 gene plays an important role in plant disease resistance signal transduction.One Pti1-like gene ZmPti1 participates in the salt resistance signal transduction,as well.To determine the subcelluar localization of ZmPti1 so as to make clearer of the ZmPti1 function,the green fluorescent protein (GFP) was used as a reporter gene to construct the plant expression vectors pGreen0029-35S ∷ ZmPti1 -GFP and pGreen002935S∷GFP.The 35S promoter was used to drive the gene expression of ZmPti1 -GFP fusion protein and pure GFP protein.Firstly,the two vectors were transformed into competent Agrobacterium strain GV3103,respectively.Then,Arabidopsis was infected by transformed Agrobacterium via floral dip method.After tansformation 62 transgenic Arabidopsis lines were got.Subcellular localization of ZmPti1 in transgenic Arabidopsis was determined by Confocal laser scanning microscopy.Results showed that ZmPti1 might be a soluble protein and localize in the cytoplasm.
分 类 号:S184[农业科学—农业基础科学]
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