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从拟南芥表达文库中筛选钙调素结合蛋白cDNA克隆

李国亮, 詹红利, 李翠凤, 刘华, 凌启阆, Zhang R

南开大学生物化学与分子生物学系.天津 300071；Department of Biological Sciences,University of Woolongong.Australia

摘要

以生物素标记的钙调素为探针,筛选拟南芥#lambda#ZAPⅡ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CaM BP-10一样,不依赖于钙离子的存在(图3A)。

关键词： 生物素标记钙调素; 钙调素结合蛋白; 表达文库筛选

Cloning of cDNAs encoding calmodulin-binding protein from Arabidopsis cDNA expression library

Li Guoliang, Zhan Hongli, Li Cuifeng, Liu Hua, Ling Qilang, Zhang R

Department of Biochemistry and Molecular Biology,Nankai University.Tianjin 300071

Abstract

Using protein-protein interaction, an Arabidopsis #lambda#ZAP II 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. Eight of them bound calmodulin depending on the presence of Ca~(2+), 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+).

Key words: Biotinylated;CaM calmodulin binding;Protein expression library screening

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引用本文：

李国亮, 詹红利, 李翠凤, 刘华, 凌启阆, Zhang R. 从拟南芥表达文库中筛选钙调素结合蛋白cDNA克隆[J]. 生理学报 2001; 53 (1): .

Li Guoliang, Zhan Hongli, Li Cuifeng, Liu Hua, Ling Qilang, Zhang R. Cloning of cDNAs encoding calmodulin-binding protein from Arabidopsis cDNA expression library. Acta Physiol Sin 2001; 53 (1): (in Chinese with English abstract).