One-step PCR mediated strategy for the construction of conditionally expressed and epitope tagged yeast proteins.
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.
Lafontaine
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European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Tollervey
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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*
D017931:DNA Primers; D000939:Epitopes; D005656:Fungal Proteins; D005819:Genetic Markers; D011993:Recombinant Fusion Proteins