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当前位置: 首页 > 文章 > 野大麦HbCBL1和HbCBL2结构与亚细胞定位分析 华北农学报 2011,26 (4) 1-7
Position: Home > Articles > Analyses for Protein Structure and Subcellular Localization of HbCBL1 and HbCBL2 from Hordeum brevisubulatum Acta Agriculturae Boreali-Sinica 2011,26 (4) 1-7

野大麦HbCBL1和HbCBL2结构与亚细胞定位分析

作  者：
吴广宇;李国婧;王瑞刚;李瑞芬;魏建华
单  位：
内蒙古农业大学生命科学学院;北京市农林科学院北京农业生物技术研究中心
关键词：
野大麦;HbCBL1;HbCBL2;激光共聚焦扫描显微镜;亚细胞定位
摘  要：
利用RT-PCR从野大麦cDNA扩增出两个基因HbCBL1和HbCBL2。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。
译  名：
Analyses for Protein Structure and Subcellular Localization of HbCBL1 and HbCBL2 from Hordeum brevisubulatum
作  者：
WU Guang-yu1,2,LI Guo-jing1,WANG Rui-gang1,LI Rui-fen2,WEI Jian-hua2(1.College of Life Science,Inner Mongolia Agricultural University,Huhhot 010018,China; 2.Beijing Agro-Biotechnology Research Center,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097,China)
关键词：
Hordeum brevisubulatum;HbCBL2;HbCBL1;Confocal laser scanning microscope(CLSM);Subcellular localization
摘  要：
RT-PCR was used to obtain two genes named as HbCBL1 and HbCBL2 from Hordeum brevisubulatum.Protein structure analysis showed that the proteins encoded by these two genes have the typical EF-hands,and HbCBL1 has N-myristoylation motif.In order to clarify whether HbCBL1 and HbCBL2 protein structures contribute to their subcellular localization,several plant expression vectors were constructed and transformed into onion epidermal cells by particle bombardment,respectively.These vectors were also transformed into Arabidopsis by floral dip.The subcellular localization of fusion proteins was determined by Confocal laser scanning microscope.The results of transient expression showed that the fusion protein HbCBL2-GFP was accumulated in tonoplast,but the HbCBL1-GFP fussion protein was accumulated in plasma membrane as well as in nucleus;N-myristoylation motif of HbCBL1 involves in its localization at cell level.The results of stable expression showed that HbCBL2 was located at tonoplast and HbCBL1 was located at plasma memebrane,additionally,both HbCBL2 and HbCBL1 are located at the guard cells.It was suggested that HbCBL1 and HbCBL2 protein structures involved in their subcellular localization,which play roles for further analyses of gene functions.

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