SUMO as a solubility tag and in vivo cleavage of SUMO fusion proteins with Ulp1.
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.
Mesh Terms:
Chromatography, Affinity, Cloning, Molecular, Cysteine Endopeptidases, Escherichia coli, Genetic Vectors, Molecular Biology, Protein Binding, Protein Biosynthesis, Recombinant Fusion Proteins, Small Ubiquitin-Related Modifier Proteins, Solubility
Chromatography, Affinity, Cloning, Molecular, Cysteine Endopeptidases, Escherichia coli, Genetic Vectors, Molecular Biology, Protein Binding, Protein Biosynthesis, Recombinant Fusion Proteins, Small Ubiquitin-Related Modifier Proteins, Solubility
Methods Mol. Biol.
Date: Jun. 20, 2014
PubMed ID: 24943315
View in: Pubmed Google Scholar
Download Curated Data For This Publication
203144
Switch View:
- Interactions 1