Protein Expression Facility, The University of Queensland, Brisbane, QLD, Australia.
Methods Mol Biol. 2022;2406:93-111. doi: 10.1007/978-1-0716-1859-2_5.
Escherichia coli remains a traditional and widely used host organism for recombinant protein production. Its well-studied genome, availability of vectors and strains, cheap and relatively straight-forward cultivation methods paired with reported high protein yields are reasons why E. coli is often the first-choice host expression system for recombinant protein production. The chapter enclosed here details protocols and design strategies in strain selection and methods on how to parallelize expression conditions to optimize for soluble target protein expression in E. coli. The methods described have been validated in a protein production research facility.
大肠杆菌仍然是传统的、广泛使用的重组蛋白生产宿主。其基因组研究透彻、载体和菌株可用性高、培养方法相对简单且成本低廉，加上报告的高蛋白质产量，这些都是大肠杆菌成为重组蛋白生产首选宿主表达系统的原因。本章详细介绍了菌株选择的方案和设计策略，以及如何并行表达条件以优化大肠杆菌中目标可溶性蛋白表达的方法。这些方法已在蛋白质生产研究机构中得到验证。
