title: Preparation of Saccharomyces cerevisiae expression plasmids.
abstract: Expression plasmids for Saccharomyces cerevisiae offer a wide choice of vector copy number, promoters of varying strength and selection markers. These expression plasmids are usually shuttle vectors that can be propagated both in yeast and bacteria, making them useful in gene cloning. For heterologous production of membrane proteins, we used the green fluorescent protein (GFP) fusion technology which was previously developed in the Escherichia coli system. We designed an expression plasmid carrying an inducible GAL1 promoter, a gene encoding a membrane protein of interest and the GFP-octa-histidine sequence. Here we describe construction of multi-copy yeast expression plasmids by homologous recombination in S. cerevisiae.
authors.affiliation: Division of Molecular Biosciences, Membrane Protein Crystallography Group, Imperial College, London, UK.
authors.forename: David
authors.initials: D
authors.lastname: Drew
authors.forename: Hyun
authors.initials: H
authors.lastname: Kim
mesh_terms: D005686:Galactokinase / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; C524219:GAL1 protein, S cerevisiae; D005686:Galactokinase