拟南芥蛋白激酶CDPK11融合蛋白的纯化及鉴定  

Recombinant Purification and Identification of Protein Kinase CDPK11 of Arabidopsis thaliana

作　　者：袁敏 邢继红[2] 王莉 张岚 葛伟娜 张家琦 

机构地区：[1]华北理工大学生命科学学院基因组学与计算生物学研究中心,河北唐山063000 [2]河北农业大学真菌毒素与植物分子病理学实验室,河北保定071001

出　　处：《河北师范大学学报（自然科学版）》2017年第6期528-532,共5页Journal of Hebei Normal University：Natural Science

基　　金：国家自然科学基金(31401212)

摘　　要：CDPK11是钙依赖蛋白激酶家族的重要成员,为了获得纯化的His-CDPK11融合蛋白,首先克隆了1 488bp的CDPK11全长基因,酶切连接构建到pGEX-4T-3载体上,获得His-CDPK11原核表达载体并转化大肠杆菌表达菌株BL21;经IPTG诱导和His镍柱体系纯化获得分子量为50ku的His-CDPK11融合蛋白;Western blot免疫印迹技术检测证实该蛋白能被Anti-His抗体识别,表明获得了正确融合的His-CDPK11蛋白,为进一步解析CDPK11参与植物开花调控的分子机制提供了有力的工具.CDPK11 is an important member of Ca2＋ dependent kinases.In order to obtain His-CDPK11 recombinant protein,the full length of 1 488 bp CDPK11 gene was cloned,digested and constructed into pGEX-4 T-3 vector to obtain His-CDPK11 prokaryotic expression construct.The recombinant His-CDPK11 plasmid was transformed into Escherichia coli strain BL21.The 50 ku recombinant His-CDPK11 protein was expressed induction with IPTG and purified by Ni-NTA affinity column.The purified protein could be detected by Anti-His antibody in western blot assay,indicating that the purified protein was correct recombinant His-CDPK11 protein.This would provide a powerful tool for further illustration of the molecular mechanism of CDPK11 mediated flowering in plants.

关 键 词：CDPK11 原核表达 融合蛋白 免疫印迹 

分 类 号：Q71[生物学—分子生物学]