D1蛋白酶的高效毛细管电泳检测  

Detection of D1 protease by high performance capillary electrophoresis

作　　者：张艳梅[1] 陈腊月[1] 李伟国[2] 郭建军[1] 刘中来[1] 姚汉超[1] 刘艳丽[1] 祁超[1] 

机构地区：[1]华中师范大学生命科学学院遗传调控与整合生物学湖北省重点实验室,430079 [2]华中师范大学化学学院,武汉430079

出　　处：《华中师范大学学报（自然科学版）》2010年第3期455-459,462,共6页Journal of Central China Normal University：Natural Sciences

基　　金：国家自然科学基金项目(20702017);教育部科技重点项目(107082);遗传调控与整合生物学湖北省重点实验室开放项目

摘　　要：在低温条件下表达的两种融合蛋白pET-28a ctpA与pET-23a-ctpA通过亲和色谱纯化、层析柱脱盐及浓缩,获得大量较高纯度的可溶性融合蛋白;并用高效毛细管电泳(HPCE)技术对两种融合蛋白进行分离检测,同时用荧光法配合确证.高效毛细管电泳分离检测结果显示,运行缓冲液的pH值相同时,两种融合蛋白的迁移时间存在显著差异;运行缓冲液的pH值在一定范围内变化时,同种蛋白的迁移时间无显著变化,表明蛋白结构是影响蛋白迁移的主要因素,两种蛋白的结构存在差异.The fusion protein pET-28a-ctpA and pET-23a-ctpA were expressed under low temperature, the expressed proteins were purified with affinity chromatography, desalted with Histrap FF and concentrated. The fusion proteins were detection by high performance capillary electrophoresis, a fluorospectrophotometer was employed to determine the structure of the reaction products at the same time to confirm the observa- tion results obtained by CZE. HPCE analysis showed that the migrate time of two fusion protein is different in the same pH of running buffer, the migrate time of one fusion protein was not significant difference when the pH of running buffer is changed with some range. It indicated that the main effect factor of migrate time is structure and the structure of two fusion protein is different.

关 键 词：D1蛋白酶 融合蛋白 高效毛细管电泳 

分 类 号：O657.8[理学—分析化学]