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Vectors for Cu2+-inducible production of glutathione S-transferase-fusion proteins for single-step purification from yeast
Authors:Alister C. Ward  Laura A. Castelli  Ian G. Macreadie  Ahmed A. Azad
Abstract:
We describe six new yeast episomal vectors which encode glutathione S-transferase (GST) affinity tags. These allow for the production of GST-fusion proteins in Saccharomyces cerevisiae under the control of the CUP1 promoter. Affinity chromatography with glutathione-Sepharose permits convenient purification of the fusion protein from a yeast lysate. The presence of a protease cleavage site facilitates subsequent removal of the GST tag. The expression and single-step purification of both GST and a functional GST-metallothionein fusion from yeast are shown as an example of the application of these vectors.
Keywords:Saccharomyces cerevisiae  CUP1  Cu2+-regulated  yeast expression  affinity purification  glutathione S-transferase  metallothionein
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