从拟南芥表达文库中筛选钙调素结合蛋白cDNA克隆  

Cloning of cDNAs Encoding Calmodulin binding Protein from Arabidopsis cDNA Expression Library

作　　者：李国亮[1] 詹红利[1] 李翠凤[1] 刘华[1] 凌启阆[1] Ren Zhng 

机构地区：[1]南开大学生物化学与分子生物学系，天津 300071 [2]Department of Biological Sciences, University of Wollongong , Australia

出　　处：《植物生理学报（0257-4829)》2001年第1期28-32,共5页Acta Phytophysiologica Sinica

基　　金：国家自然科学基金委资助项目 !(批准号 :396 70 16 4)

摘　　要：以生物素标记的钙调素为探针 ,筛选拟南芥λZAPⅡ表达文库 ,分离得到 9个阳性克隆。融合蛋白的Western印迹分析表明 ,这些阳性克隆确是编码钙调素结合蛋白的 (图 2 ,3) ,并且其中Y9等 8个克隆编码的融合蛋白与钙调素有依赖于钙离子的结合 (图3B) ;而唯独Y7编码的融合蛋白与钙调素的结合 ,与CaMBP 10一样 ,不依赖于钙离子的存在 (图 3A)。Using protein protein interaction, an Arabidopsis λZAPⅡ cDNA expression library was screened with biotinylated calmodulin as a probe. Nine positive clones were isolated. The results of Western blotting of fusion proteins indicated that all these positive clones could synthesize peptides which could bind calmodulin (Figs.2,3). Eight of them bound calmodulin depending on the presence of Ca 2+ (Fig.3B), but only one fusion protein encoded by Y7 cDNA clone, which was the same as CaM BP 10, could bind CaM in a way independent of Ca 2+ (Fig.3A).

关 键 词：生物素标 钙调素结合蛋白 表达文库筛选 克隆 

分 类 号：Q943.2[生物学—植物学]