A novel self-cleavage system for production of soluble recombinant protein in Escherichia coli

文献类型： 外文期刊

第一作者： Feng, Yufei

作者： Feng, Yufei;Wu, Donglai;Feng, Yufei;Xu, Qingyuan;Yang, Tao;Sun, Encheng;Li, Junping;Wu, Donglai;Shi, Dongfang

作者机构：

关键词： Self-cleavage;Protein expression;Human rhinovirus 3C protease (HRV3C);Escherichia coli (E. coli)

期刊名称：PROTEIN EXPRESSION AND PURIFICATION （ 影响因子：1.65； 五年影响因子：1.548 ）

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收录情况： SCI

摘要： Many approaches for generating large quantities of recombinant protein in Escherichia coli fuse the protein of interest to a protein tag to enhance solubility and improve recovery. However, the fusion tags can confound downstream applications, as the fusion partner can alter the structure and biological activity of the recombinant protein and proteolytic removal of the fusion tags can be expensive. Here we describe a new system for production of native proteins in E. coli that allows for removal of the fusion tag via intracellular self-cleavage by the human rhinovirus 3C (HRV3C) protease. This system allows for parallel cloning of target protein coding sequences into six different expression vectors, each with a different fusion partner tag to enhance solubility during induction. Temperature-regulated expression of the HRV3C protease allows for intracellular removal of the fusion tag following induction, and the liberated recombinant protein can be purified by affinity chromatography by virtue of a short six-histidine tag. This system will be an attractive approach for the expression and purification of recombinant proteins free of solubilityenhancing fusion tags, and should be amenable to high-throughput applications.

分类号： Q51

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