Recombinant Protein Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay.
Biotechnol J. 2011 Jun;6(6):715-30. doi: 10.1002/biot.201100025. Epub 2011 May 12.
Proteins are the main reagents for structural, biomedical, and biotechnological studies; however, some important challenges remain concerning protein solubility and stability. Numerous strategies have been developed, with some success, to mitigate these challenges, but a universal strategy is still elusive. Currently, researchers face a plethora of alternatives for the expression of the target protein, which generates a great diversity of conditions to be evaluated. Among these, different promoter strength, diverse expression host and constructs, or special culture conditions have an important role in protein solubility. With the arrival of automated high-throughput screening (HTS) systems, the evaluation of hundreds of different conditions within reasonable cost and time limits is possible. This technology increases the chances to obtain the target protein in a pure, soluble, and stable state. This review focuses on some of the most commonly used strategies for the expression of recombinant proteins in the enterobacterium Escherichia coli, including the use of HTS for the production of soluble proteins.
蛋白质是结构、生物医学和生物技术研究的主要试剂；然而，关于蛋白质的溶解度和稳定性仍然存在一些重要的挑战。已经开发了许多策略来减轻这些挑战，但是仍然缺乏一种通用的策略。目前，研究人员面临着大量表达目标蛋白的替代方案，这产生了大量需要评估的条件。在这些条件中，不同的启动子强度、不同的表达宿主和构建体或特殊的培养条件对蛋白质的溶解度有重要作用。随着自动化高通量筛选（HTS）系统的出现，在合理的成本和时间限制内评估数百种不同的条件成为可能。该技术增加了以纯、可溶性和稳定状态获得目标蛋白的机会。这篇综述集中讨论了在大肠杆菌中表达重组蛋白的一些最常用的策略，包括使用 HTS 生产可溶性蛋白。
