Loughran Sinéad T, Bree Ronan T, Walls Dermot
Department of Applied Sciences, School of Health and Science, Dundalk Institute of Technology, Dundalk, Louth, Ireland.
School of Biotechnology, Dublin City University, Dublin 9, Ireland.
Methods Mol Biol. 2017;1485:275-303. doi: 10.1007/978-1-4939-6412-3_14.
His-tagging is the most widespread and versatile strategy used to purify recombinant proteins for biochemical and structural studies. Recombinant DNA methods are first used to engineer the addition of a short tract of poly-histidine tag (His-tag) to the N-terminus or C-terminus of a target protein. The His-tag is then exploited to enable purification of the "tagged" protein by Immobilized Metal Affinity Chromatography (IMAC). Here, we describe efficient procedures for the isolation of highly purified His-tagged target proteins from an E. coli host using IMAC.
His标签法是用于纯化重组蛋白以进行生化和结构研究的最广泛且通用的策略。首先使用重组DNA方法在目标蛋白的N端或C端设计添加一小段多组氨酸标签（His标签）。然后利用His标签通过固定化金属亲和色谱法（IMAC）纯化“带标签”的蛋白。在此，我们描述了使用IMAC从大肠杆菌宿主中高效分离高纯度His标签化目标蛋白的方法。
