title: One-step PCR mediated strategy for the construction of conditionally expressed and epitope tagged yeast proteins.
abstract: With the availability of the complete yeast genomic sequence, techniques which allow the rapid functional analysis of genes of interest are of increasing importance. Here we report a technique which allows the initial characterisation of genes of interest, through the construction of conditionally expressed mutations for functional analyses and the generation of epitope-tagged fusion proteins for immuno-localisation and immuno-purification, entirely by PCR.
authors.affiliation: European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Lafontaine
authors.forename: D
authors.initials: D
authors.lastname: Tollervey
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D000939:Epitopes; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005819:Genetic Markers; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D017931:DNA Primers; D000939:Epitopes; D005656:Fungal Proteins; D005819:Genetic Markers; D011993:Recombinant Fusion Proteins