Difference between revisions of "Part:BBa K3351009"

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His tag.

Summary

A major time-consuming process in nearly all structural and functional studies of proteins is their overproduction and purification. Recombinant protein production in Escherichia coli has become the most popular platform for researchers who require large amounts of protein. Immobilized metal affinity chromatography (IMAC) with a polyhistidine tag (usually six consecutive histidine residues) has emerged as the most common and convenient method for purifying recombinant proteins. His tag has chelations of a variety of metal ions, including Ca2+, Mg2+, Ni2+, Co2+, etc, among which nickel ions are most widely used. His tags have gained great popularity over the last decade as a purification tool for recombinant proteins. Cloning vectors generally introduce six consecutive histidines and an optional protease-cleavage site or linker to the N- or C-terminus of the protein of interest. These His tags facilitate selective binding of the expressed protein to a nickel-affinity column. The tag may then optionally be removed by a protease, requiring another purification step.

Reference

[1] Smith, M. C., Furman, T. C., Ingolia, T. D. & Pidgeon, C. (1988). J. Biol. Chem. 263, 7211–7215 [2] Carson M , Johnson D H , Mcdonald H , et al. His-tag impact on structure[J]. Acta Crystallographica, 2007, 63(3):295-301. [3] Raran-Kurussi S, Waugh DS. Expression and Purification of Recombinant Proteins in Escherichia coli with a His6 or Dual His6-MBP Tag. Methods Mol Biol. 2017;1607:1-15. doi: 10.1007/978-1-4939-7000-1_1. PMID: 28573567; PMCID: PMC7122414. Sequence and Features