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Targeted expression, purification, and cleavage of fusion proteins from inclusion bodies in Escherichia coli
Author(s) -
Hwang Peter M.,
Pan Jonathan S.,
Sykes Brian D.
Publication year - 2014
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2013.09.028
Subject(s) - cyanogen bromide , fusion protein , inclusion bodies , chemistry , escherichia coli , cleavage (geology) , ketosteroid , biochemistry , fusion , formic acid , isomerase , combinatorial chemistry , peptide sequence , biology , enzyme , paleontology , fracture (geology) , gene , recombinant dna , linguistics , philosophy
There are many fusion protein systems now available for insoluble expression: TrpLE, ketosteroid isomerase, PurF, and PagP, for example. The ideal fusion partner is effective at directing a wide variety of target proteins into inclusion bodies, accumulates in large quantities in a highly pure form, and is readily solubilized and purified in commonly used denaturants. Fusion partner removal under denaturing conditions is biochemically challenging, requiring harsh conditions (e.g., cyanogen bromide in 70% formic acid) that can result in unwanted protein modifications. Recent advances in metal ion‐catalyzed peptide bond cleavage allow for more mild conditions, and some methods involving nickel or palladium will likely soon appear in more biological applications.