Evaluation of a Cold Shock Promoter System for the Production of Recombinant Proteins in E. coli

用于在大肠杆菌中生产重组蛋白的冷激启动子系统的评估

• 批准号: 9308294
• 负责人: Francois Baneyx
• 金额: $ 9.91万
• 依托单位: University of Washington
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-06-15 至 1996-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9308294&HistoricalAwards=false
• 关键词: Evaluation; Cold; Shock; Promoter; System

The focus of this project is to evaluate the potential of cold shock promoters for the expression of recombinant proteins in Escherichia coli following a shift to a low temperature culturing condition. The primary objective of this project is to characterize the expression of different model recombinant proteins placed under the control of the cspA cold shock promoter. The strength and inducibility of the cold shock promoter in directing the low temperature synthesis of proteins which are produced in a biologically active conformation under normal growth conditions will be first compared and contrasted to those of the widely used tac promoter. Potential advantages of cold shock promoters are facilitated folding, and a reduced degradation of recombinant proteins. The cspA-controlled production of a recombinant protein susceptible to aggregation, and a recominant protein susceptible to degradation under normal growth conditions will be specifically investigated. The secondary objective of this project is to extend initial shake flask studies to pilot scale fermentations in order to demonstrate the usefulness of cold shock promoters for the large-scale production of useful proteins.

该项目的重点是 评估冷休克的可能性 启动子的表达 大肠杆菌重组蛋白 大肠杆菌后转移到低 温度培养条件 这项工作的主要目的是 该项目旨在描述 不同模型表达 重组蛋白置于 控制cspA冷休克 启动子 强度和 冷休克诱发 启动子在指导低 蛋白质的温度合成 它们是在生物学上 正常下活性构象 生长条件将是第一 与那些 广泛使用的tac启动子。 冷休克的潜在优势 启动子易于折叠， 和降低的降解， 重组蛋白 的 cspA控制的生产 重组蛋白易感 聚合，和一个建议 易降解蛋白质 在正常生长条件下， 具体调查。 的 本项目的次要目标 扩大初始摇瓶 中试规模发酵研究 为了证明 冷休克促进剂的用途 规模化生产 有用的蛋白质。