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

本项目的重点是评估低温培养条件下冷休克启动子在大肠杆菌中重组蛋白表达的潜力。本项目的主要目的是表征在cspA冷休克启动子控制下不同模型重组蛋白的表达。首先是冷休克启动子的强度和诱导作用，以指导在正常生长条件下以生物活性构象产生的蛋白质的低温合成与广泛应用的tac启动子进行了比较。冷休克启动子的潜在优势是易于折叠和减少重组蛋白的降解。在正常生长条件下，将特别研究cspa控制的易聚集的重组蛋白和易降解的重组蛋白的生产。该项目的第二个目标是将最初的摇瓶研究扩展到中试规模发酵，以证明冷冲击促进剂对大规模生产的有用性有用的蛋白质。

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.