Sugiki Toshihiko, Fujiwara Toshimichi, Kojima Chojiro
Graduate School of Engineering, Yokohama National University, Yokohama, Japan.
Methods Mol Biol. 2017;1586:345-357. doi: 10.1007/978-1-4939-6887-9_23.
The cold-shock system using the pCold vector is one of the most effective Escherichia coli heterologous protein expression systems. It allows the improvement of the expression level of the protein of interest in a soluble fraction. In this chapter, we describe practical procedures for the overexpression of heterologous protein of interest by using the pCold vector or the single-protein production system. The latter is one of the most advanced pCold technologies for isotope labeling of the target protein and its NMR studies.
使用pCold载体的冷休克系统是最有效的大肠杆菌异源蛋白表达系统之一。它能够提高目的蛋白在可溶部分的表达水平。在本章中，我们描述了使用pCold载体或单蛋白生产系统过表达目的异源蛋白的实际操作步骤。后者是用于目标蛋白同位素标记及其核磁共振研究的最先进的pCold技术之一。
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