一种新的分子内可自剪切多肽介导的可视化原核蛋白质表达系统  

A Visualized Prokaryotic Protein Expression System Mediated by a New Intramolecular Self-cleavage Peptide

作　　者：吴惠惠 黄浩[1] 易敏 杨爱芬 杜君卿 WU Huihui;HUANG Hao;YI Min;YANG Aifen;DU Junqing(School of Life and Environmental Sciences,Hangzhou Normal University,Hangzhou 311121,China)

机构地区：[1]杭州师范大学生命与环境科学学院

出　　处：《杭州师范大学学报（自然科学版）》2019年第5期499-503,共5页Journal of Hangzhou Normal University(Natural Science Edition)

基　　金：国家自然科学基金项目(31771621);浙江省自然科学青年基金项目(Q16C090004,LQ19C090001)

摘　　要：通过在目标蛋白和荧光蛋白等标签蛋白之间引入可引起分子内剪切的多肽是监测蛋白质表达的重要方法.通过将小鼠MYRF的介导其分子内自剪切的ICA结构域插入到目标蛋白和指示蛋白mCherry之间,既实现了对融合蛋白表达的直接观察,同时实现了融合蛋白的高效自剪切,并释放出游离的目标蛋白.实验证明,该可视化蛋白质表达系统可在原核细胞中有效的表达不携带标签序列的目标蛋白.Intramolecular self-cleavage peptides fused with visual tags,such as fluorescent proteins,are widely used in illuminating the expression of the target proteins.By inserting the intramolecular chaperone auto-processed domain(ICA)of mouse transcriptional factor MYRF between target proteins and a fluorescent protein mCherry,the expression of target proteins can be detected visually,and the free target proteins can be released from fusion proteins efficiently.The data show that the visualized protein expression system mediated by ICA self-cleavage domain is capable of supporting tag-free protein expression in prokaryotic cells.

关 键 词：MYRF ICA结构域 分子内自剪切 原核表达 可视化 

分 类 号：Q816[生物学—生物工程]