Vectors for Cu2+-inducible production of glutathione S-transferase-fusion proteins for single-step purification from yeast |
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Authors: | Alister C. Ward Laura A. Castelli Ian G. Macreadie Ahmed A. Azad |
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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. |
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Keywords: | Saccharomyces cerevisiae CUP1 Cu2+-regulated yeast expression affinity purification glutathione S-transferase metallothionein |
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