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Vectors for Cu2+-inducible production of glutathione S-transferase-fusion proteins for single-step purification from yeast
journal contribution
posted on 1994-04-01, 00:00 authored by Alister WardAlister Ward, L A Castelli, I G Macreadie, A A AzadWe 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.
History
Journal
YeastVolume
10Issue
4Pagination
441 - 449Publisher
John Wiley & SonsLocation
Chichester, Eng.Publisher DOI
ISSN
0749-503XLanguage
engPublication classification
C1.1 Refereed article in a scholarly journalCopyright notice
1994, John Wiley & SonsUsage metrics
Categories
Keywords
Amino Acid SequenceAnimalsBase SequenceChromatography, AffinityCopperEnzyme InductionEscherichia coliFactor XaGene Expression Regulation, FungalGenetic VectorsGlutathione TransferaseMetallothioneinMolecular Sequence DataPromoter Regions, GeneticProtozoan ProteinsRecombinant Fusion ProteinsSaccharomyces cerevisiaeSchistosoma japonicumThrombinCUP1Cu2+yeast expressionaffinity puriticationglutathioneS-transferaseScience & TechnologyLife Sciences & BiomedicineBiochemistry & Molecular BiologyBiotechnology & Applied MicrobiologyMicrobiologyMycologyCU2+-REGULATEDAFFINITY PURIFICATIONGLUTATHIONE S-TRANSFERASEESCHERICHIA-COLISHUTTLE VECTORSTRANSFORMATIONEXPRESSIONCLEAVAGE