大肠杆菌热休克或冷休克启动子调控的基因表达载体

The vectors using heat or cold shock promoter in Escherichia coli to control gene expression

摘要:

基因重组技术已经成为获得各种酶和生物活性蛋白的主要手段。虽然很多基因已在大肠杆菌中得到高效表达，但是当人们认定某种蛋白对科学研究或生产应用极为重要时，却常常因为其基因表达水平很低或产生包涵体而感到束手无策。表达载体pHsh和pEXC通过激活热休克或冷休克转录调控机制提高分子伴侣的表达水平，从而降低目标蛋白的细胞毒性并减少包涵体形成。应用于生物合成、分子修饰或生物降解的高温酶可以通过pHsh系统表达获得高产，而科研和诊疗所需要的来源于动植物和常温微生物的基因可以通过pEXC系统获得高效表达。这些新载体的发展为重组蛋白的小规模制备和大规模生产提供了新策略和有效途径。

Abstract:

Gene recombination technology has become a main route to produce proteins in laboratory and industry. Though many proteins have been efficiently expressed in Escherichia coli, people have difficulties when target genes express at a very low level or produce inclusion body, especially for the proteins important for research and production. The expression level of molecular chaperones can be enhanced by the transcription factors of a heat shock or cold shock system in vectors pHsh and pEXC, which could reduce cytotoxicity of target proteins and formation of inclusion body. Thermostable enzymes used in biosynthesis, molecular modification or biodegradation could be overexpressed in pHsh expression system, while the genes from animals, plants and mesophilic microorganisms can be effectively expressed by using vector pEXC. The development of these novel vectors supply new strategies and effective approaches for small-scale preparation and industrial production of enzymes and bioactive proteins.