标记重组蛋白以提高溶解度并辅助纯化。

Tagging Recombinant Proteins to Enhance Solubility and Aid Purification.

作者信息

Loughran Sinéad T, Walls Dermot

机构信息

Department of Applied Sciences, Dundalk Institute of Technology, Dundalk, Ireland.

School of Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland.

出版信息

Methods Mol Biol. 2017;1485:131-156. doi: 10.1007/978-1-4939-6412-3_8.

DOI:10.1007/978-1-4939-6412-3_8

PMID:27730552

Abstract

Protein fusion technology has had a major impact on the efficient production and purification of individual recombinant proteins. The use of genetically engineered affinity and solubility-enhancing polypeptide "tags" has increased greatly in recent years and there now exists a considerable repertoire of these that can be used to solve issues related to the expression, stability, solubility, folding, and purification of their fusion partner. In the case of large-scale proteomic studies, the development of purification procedures tailored to individual proteins is not practicable, and affinity tags have therefore become indispensable tools for structural and functional proteomic initiatives that involve the expression of many proteins in parallel. Here, the rationale and applications of a range of established and more recently developed solubility-enhancing and affinity tags is described.

摘要

蛋白质融合技术对单个重组蛋白的高效生产和纯化产生了重大影响。近年来，基因工程亲和性和溶解性增强多肽“标签”的使用大幅增加，目前已有相当多的此类标签可用于解决与其融合伙伴的表达、稳定性、溶解性、折叠和纯化相关的问题。在大规模蛋白质组学研究中，针对单个蛋白质量身定制纯化程序是不可行的，因此亲和标签已成为结构和功能蛋白质组学计划中不可或缺的工具，这些计划涉及同时表达多种蛋白质。本文描述了一系列已确立的和最近开发的溶解性增强标签及亲和标签的原理和应用。