使用自动加工的可诱导酶标签简化蛋白质纯化。
Simplified protein purification using an autoprocessing, inducible enzyme tag.
作者信息
Shen Aimee
机构信息
Department of Microbiology and Molecular Genetics, University of Vermont, 95 Carrigan Drive, Burlington, VT, 05405, USA,
出版信息
Methods Mol Biol. 2014;1177:59-70. doi: 10.1007/978-1-4939-1034-2_5.
The development of affinity tags has greatly simplified protein purification procedures. A variety of affinity tags are now available to improve expression, solubility, and/or tag removal. In this chapter, we describe a method for purifying recombinant proteins expressed in Escherichia coli that uses a highly specific, inducible, C-terminal autoprocessing protease tag. This method streamlines affinity purification, cleavage, and tag separation into a one-step purification procedure, avoiding the need to remove fusion tags from target proteins with exogenous proteases. In addition to accelerating protein purification, we show that this method can enhance the expression, stability, and solubility of select proteins.
亲和标签的发展极大地简化了蛋白质纯化程序。现在有多种亲和标签可用于提高表达、溶解性和/或去除标签。在本章中，我们描述了一种纯化在大肠杆菌中表达的重组蛋白的方法，该方法使用一种高度特异性、可诱导的C端自切割蛋白酶标签。这种方法将亲和纯化、切割和标签分离简化为一步纯化程序，避免了使用外源蛋白酶从目标蛋白上去除融合标签的需要。除了加速蛋白质纯化外，我们还表明这种方法可以提高某些蛋白质的表达、稳定性和溶解性。
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