一种新的烟草钙调素结合蛋白的分离鉴定

一种新的烟草钙调素结合蛋白的分离鉴定 Isolation and Characterization of a cDNA Encoding a New Calmodulin-Binding Protein from Tobacco 王芸 1 2 3 4 first-author 杨万年 1 2 3 4 吕应堂 1 2 3 4 武汉大学生命科学学院植物发育生物学教育部重点实验室 武汉大学生命科学学院植物发育生物学教育部重点实验室 华中师范大学生命科学学院 华中师范大学生命科学学院 ,湖北,武汉,430079 ,湖北,武汉,430079 用35S标记的钙调素（35S CaM）作探针,从烟草cDNA文库中筛选到一个cDNA克隆pTCB40.DNA测序表明,分离的cDNA片段具有的开放阅读框编码233个氨基酸的多肽链.所编码钙调素结合蛋白TCB40（CaM bindingprotein,CaMBP）与离子通道蛋白的同源性高达81%.凝胶覆盖实验结果证明其表达的蛋白具有依赖Ca2+的钙调素结合活性.缺失分析表明钙调素结合位点于其蛋白的C端.Northernblot分析发现,TCB40在烟草茎,叶和花中均表达,但在叶中的表达量明显高于其他器官. A cDNA clone encoding a new calmodulin-binding protein （TCB40） was isolated by screening cDNA expression library with 35 S-labled CaM. DNA sequence analysis indicated that this cDNA have one open reading frame encoding a peptide of 233 amino acid residues for TCB40 that share high homology with ion channel. Gel overlay experiments demonstrated that TCB40 bound to CaM in a calcium-dependent manner. By molecular dissection analyses, the calmodulin-binding domain of this protein was localized at C-terminal of TCB40. Northern blot indicated that TCB40 gene is expressed in stem, leaf and flower of tobacco, with high abundance in leaf. 钙调素结合蛋白 凝胶覆盖分析 钙调素结合功能区 calmodulin-binding protein gel overlay experiments calmodulin-binding domain Q946 国家自然科学基金资助项目（30070078）;高校博士点基金及武汉大学植物发育生物学教育部重点实验室开放课题资助 2003-06-01 2021-04-01 6