MBP-hmcm6融合蛋白的构建与表达  

The construction and expression of MBP-hmcm6

作　　者：王蕾[1] 余波光[1] 吴文言[2] 

机构地区：[1]广州中山大学有害生物控制与资源利用国家重点实验室,广州510278 [2]广州中山大学生命科学学院

出　　处：《实用肿瘤学杂志》2005年第5期321-324,共4页Practical Oncology Journal

摘　　要：目的构建含微小染色体维持蛋白MBP-hmcm6的原核表达重组质粒,并进行可溶性表达与初步纯化。方法以常规克隆方法将目的片段连接至pMAL-p2x载体,获得的阳性克隆转化大肠杆菌Rosetta(DE3)进行表达。结果经Western blot鉴定,获得了分子量大小为139 KD的MBP-hMCM6p蛋白。结论成功构建了pMAL-p2x-hmcm6重组质粒和工程菌,实现了重组蛋白可溶性表达,有助于进一步研究MCM蛋白的结构与功能。Objective To achieve the soluble expression of human MCM6p, pMAL- p2x - hmcm6 was constructed and then recombinant hmcm6 was produced in E. coli Rosetta （DE3） as a maltose - binding protein （MBP） fusion protein using the pMAL- p2x expression system. The apparent molecular weight of hmcm6p was 139 kD according to SDS- PAGE. Western blotting showed that the recombinant protein could interact with the multi - elonal antibody of protein and the soluble MBP- hmcm6p next was purified by Amylose Resin affanity chromatography.

关 键 词：微小染色体维持蛋白 pMAL-p2x 可溶性表达 融合蛋白 Western 可溶性表达 重组质粒 克隆转化 原核表达 初步纯化 大肠杆菌 BLOT 

分 类 号：R73-3[医药卫生—肿瘤]