Vectors for Cu2+-inducible production of glutathione S-transferase-fusion proteins for single-step purification from yeast
Version 2 2024-06-03, 21:45Version 2 2024-06-03, 21:45
Version 1 2017-05-16, 15:53Version 1 2017-05-16, 15:53
journal contribution
posted on 2024-06-03, 21:45authored byAlister WardAlister Ward, LA Castelli, IG Macreadie, AA Azad
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.