Screening Fusion Tags for Improved Recombinant Protein Expression in E. coli with the Expresso® Solubility and Expression Screening System.
Screening Fusion Tags for Improved Recombinant Protein Expression in E. coli with the Expresso® Solubility and Expression Screening System.
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DOI：
10.1002/cpps.39
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发表时间：
2017-11-01
影响因子：
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通讯作者：
Auldridge ME
中科院分区：
文献类型：
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作者：
Steinmetz EJ;Auldridge ME
细菌培养的简单、快速和低成本使大肠杆菌成为大多数重组蛋白表达初始试验的选择系统。然而，许多异源蛋白要么在细菌中表达不良，要么以不正确折叠的形式产生，不溶性聚集体缺乏天然蛋白的活性。在许多情况下，与伴侣蛋白的融合可以改善困难靶蛋白的表达和/或溶解度。虽然有几种不同的融合伙伴受到青睐，但没有一种是普遍有效的，并且确定最能提高给定靶蛋白可溶性表达的融合伙伴是一个经验过程。本单元提出了一种平行筛选融合伙伴的策略，以增强表达或溶解度。Expresso溶解度和表达筛选系统包括7个不同的融合伙伴的面板，并利用一个极其简单的克隆策略，使快速筛选和鉴定最有效的融合伙伴。
The simplicity, speed, and low cost of bacterial culture make E. coli the system of choice for most initial trials of recombinant protein expression. However, many heterologous proteins are either poorly expressed in bacteria, or are produced as incorrectly folded, insoluble aggregates that lack the activity of the native protein. In many cases, fusion to a partner protein can allow for improved expression and/or solubility of a difficult target protein. Although several different fusion partners have gained favor, none is universally effective, and identifying the one that best improves soluble expression of a given target protein is an empirical process. This unit presents a strategy for parallel screening of fusion partners for enhanced expression or solubility. The Expresso Solubility and Expression Screening System includes a panel of 7 distinct fusion partners, and utilizes an extremely simple cloning strategy to enable rapid screening and identification of the most effective fusion partner.