Abstract
Using the dextran-binding domain (DBD) of a type of glucosyltransferase (GTF) from Streptococcus sobrinus, we have developed a novel method for purifying recombinant proteins. DBD-tagged green and red fluorescent proteins as well as the parent GTF and DBD moiety were adsorbed well to commercially available cross-linked dextran (such as Sephadex beads and Sephacryl beads), and eluted efficiently with water-soluble dextran. The purity of the eluted proteins after this one-step affinity purification was approximately 90% or better. The results suggest that DBD can be used as a powerful carrier for purification of various recombinant proteins.
MeSH terms
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Acrylic Resins / chemistry
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Acrylic Resins / metabolism
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Chromatography, Affinity / economics
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Chromatography, Affinity / methods
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Dextrans / chemistry
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Dextrans / metabolism*
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Glycosyltransferases / genetics
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Green Fluorescent Proteins
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Luminescent Proteins / genetics
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Molecular Weight
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Protein Binding / physiology
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Protein Structure, Tertiary / physiology
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / isolation & purification*
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification*
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Recombinant Proteins / metabolism*
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Red Fluorescent Protein
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Streptococcus sobrinus
Substances
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Acrylic Resins
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Dextrans
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Luminescent Proteins
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Recombinant Fusion Proteins
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Recombinant Proteins
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Green Fluorescent Proteins
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sephacryl S 500
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sephadex
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Glycosyltransferases