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野大麦HbCBL1和HbCBL2结构与亚细胞定位分析

DOI: 10.7668/hbnxb.2011.04.001, PP. 1-7

Keywords: 野大麦,HbCBL2,HbCBL1,激光共聚焦扫描显微镜,亚细胞定位

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Abstract:

利用RT-PCR从野大麦cDNA扩增出两个基因HbCBL2和HbCBL1。蛋白结构分析表明,其具有典型的EF手臂,且HbCBL1的N-端含有豆蔻酰化结构域。为了明确野大麦HbCBL1和HbCBL2结构对其在植物体内亚细胞水平分布的作用,分别构建了以35S为启动子驱动的与绿色荧光蛋白基因GFP融合的植物表达载体,通过基因枪介导法将重组载体转入洋葱上表皮细胞,同时通过醮花法获得拟南芥稳定转基因株系,利用激光共聚焦扫描显微镜检测融合蛋白的表达部位。瞬时表达结果表明,融合蛋白HbCBL2::GFP在液泡膜上可见;HbCBL1::GFP在质膜上可见,同时在核中也有信号;HbCBL1的N端豆蔻酰化位点参与了该基因的定位。稳定表达结果表明,HbCBL2主要定位于液泡膜上;HbCBL1主要定位于质膜和细胞核;同时这两个蛋白在保卫细胞中也有表达。研究表明,HbCBL1和Hb-CBL2的结构决定了其亚细胞水平的表达部位,为进一步分析其生物学功能奠定了基础。

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