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A Mutant Sumo Facilitates Quick Plasmid Construction for Expressing Proteins with Native N-termini After Tag Removal

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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.

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Abbreviations

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

ULP/ULP1:

Ubl-specific protease 1

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Authors and Affiliations

  1. U.S. Department of Agriculture, Agricultural Research Service, Pacific West Area, Western Regional Research Center, 800 Buchanan Street, Albany, CA, 94710, USA

    Yuzhu Zhang & Yuting Fan

  2. School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China

    Yuting Fan

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  1. Yuzhu Zhang
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  2. Yuting Fan
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Correspondence to Yuzhu Zhang.

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Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and employer.

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Zhang, Y., Fan, Y. A Mutant Sumo Facilitates Quick Plasmid Construction for Expressing Proteins with Native N-termini After Tag Removal. Mol Biotechnol 59, 159–167 (2017). https://doi.org/10.1007/s12033-017-9998-6

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  • DOI: https://doi.org/10.1007/s12033-017-9998-6

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