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SUMO as a Solubility Tag and In Vivo Cleavage of SUMO Fusion Proteins with Ulp1

  • Dennis Kuo
  • Minghua Nie
  • Albert J. CoureyEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1177)

Abstract

Expression of proteins in E. coli is often plagued by insolubility of the protein of interest. A solution to this problem is the expression of proteins as fusions to solubility tags such as the SUMO protein. SUMO fusion proteins can be cleaved to remove the SUMO moiety using SUMO-specific proteases such as Ulp1. Here, we describe the use of vectors for the expression of recombinant proteins in E. coli as fusions to the Drosophila SUMO protein. This includes a vector that encodes not only the SUMO tagged protein of interest but also SUMO-tagged Ulp1. Coexpression of these two proteins results in the in vivo cleavage of the protein of interest from the SUMO tag, while still leaving the protein of interest in a form that can be purified from a soluble cell lysate by nickel affinity chromatography.

Key words

SUMO Ulp1 Solubility tag Ni-NTA resin Affinity chromatography 

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Chemistry & Biochemistry, Molecular Biology InstituteUniversity of CaliforniaLos AngelesUSA

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