Proteases and tags for protein purification and analysis.

描述(申请人提供)：这个项目的总体目标是创建一个从克隆基因中表达、检测和快速纯化蛋白质的集成系统。这个系统的基础是一种高度工程的枯草杆菌酶，它能在化学触发的情况下对底物进行水解性反应。枯草杆菌蛋白原结构域(PROTAG)将被开发用于检测天然目的蛋白，并作为亲和纯化的配基。工程加工枯草杆菌蛋白(PSUB)既可作为亲和纯化融合蛋白的结合分子，又可用作去除PROTAG的加工酶。长期目标是开发一套复杂的工具来促进蛋白质组分析，就像限制性内切酶用于促进基因组分析一样。

DESCRIPTION (provided by applicant): The overall goal to this project is to create an integrated system for the expression, detection and rapid purification of proteins from cloned genes. The foundation for this system is a highly engineered subtilisin which hydrolyzes substrates in response to a chemical trigger. The subtilisin prodomain (protag) will be developed both for detection of native target protein and as the ligand for affinity purification. The engineered processing subtilisin (psub) will be used both as the binding molecule for affinity purification of protagged fusion proteins and as the processing protease for protag removal. The long range objective is to develop a sophisticated set of tools to facilitate proteomic analysis in the way restriction endonucleases have been used to facilitate genomic analysis.