title: A Mutant Sumo Facilitates Quick Plasmid Construction for Expressing Proteins with Native N-termini After Tag Removal.
abstract: Sumo is one of the fusion tags commonly used to enhance the expression and the solubility of recombinant proteins. One advantage of using sumo is that the removal of the sumo tag is highly specific because its recognition by a sumo protease is determined by its structural characteristics, instead of the sequence of a short peptide. Recently, it was reported that sumo could also be used as a protease recognition site to facilitate the removal of other fusion tags, such as MBP, when sumo itself is not suitable to enhance the solubility of a particular target protein. Using sumo as a recognition site is highly desirable when the target protein needs to have its native N terminus. However, constructing such a plasmid involves more than one cloning step because the N terminus of the target protein needs to be the next residue after the diglycine of sumo. Here, we report the construction of a new vector with a mutant sumo tag. The incorporation of a Pvu II site near the 3' end of tag coding sequence enables quick construction of plasmids for producing proteins with native termini. Its usage includes producing recombinant food allergens for studying conformational IgE epitopes.
authors.affiliation: U.S. Department of Agriculture, Agricultural Research Service, Pacific West Area, Western Regional Research Center, 800 Buchanan Street, Albany, CA, 94710, USA. Yuzhu.Zhang@ars.usda.gov.
authors.forename: Yuzhu
authors.identifier: http://orcid.org/0000-0001-7882-5692
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: U.S. Department of Agriculture, Agricultural Research Service, Pacific West Area, Western Regional Research Center, 800 Buchanan Street, Albany, CA, 94710, USA.|School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.
authors.forename: Yuting
authors.initials: Y
authors.lastname: Fan
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D020224:Expressed Sequence Tags / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D025842:SUMO-1 Protein / Q000235:genetics*
chemical_list: D011993:Recombinant Fusion Proteins; D025842:SUMO-1 Protein
keywords: Sumo; Tandem affinity purification; ULP; pTS1