通过断裂内含肽介导的反式剪接合成大的蛋白  

Large Protein Production Through Split Intein-Mediated Trans-Splicing

作　　者：张静[1] 刘环[1] 周晶[1] 刘建华[1] 

机构地区：[1]上海交通大学生命科学技术学院,上海200240

出　　处：《中国生物工程杂志》2009年第12期74-78,共5页China Biotechnology

基　　金：国家"973"计划子课题(2007CB914500)资助项目

摘　　要：大肠杆菌难以表达大的蛋白,毒性蛋白以及膜蛋白,"Npu DnaE内含肽表达系统"使这些蛋白的表达成为可能。该系统的基本原理是:在特定位点处将目标基因(编码T7 RNA聚合酶的基因)断裂成两部分,然后分别与Npu DnaE内含肽的N端,C端片段融合,两种融合基因分别表达纯化,在体外将两种融合蛋白等摩尔比混合即可产生有功能的T7 RNA聚合酶。理论上,该体系也可用于合成其他大的蛋白,毒性蛋白或膜蛋白。Npu DnaE intein was used to produce some large proteins, which were difficult to obtain through conventional expression systems. A T7 expression system was described, by which the gene of T7 RNA polymerase is split into two pieces, and each piece fuses with Npu DnaE N- and C- terminal sequences respectively. Functional T7 RNA polymerase is created by mixing the two kinds of fusion constructs in vitro. The approach of split intein-mediated production of large proteins, in theory, readily generalizable to the purification of other large, cytotoxic or membrane proteins.

关 键 词：大的蛋白 毒性蛋白 膜蛋白 NPU DnaE内含肽 T7 RNA聚合酶 

分 类 号：Q819[生物学—生物工程]