Constitutive promoter modules for PCR-based gene modification in Saccharomyces cerevisiae.
Initial steps in investigating gene function often include deleting and overexpressing the gene of interest and identifying the subcellular location of the gene product. To facilitate these procedures, a number of new PCR modules, which contain selectable markers and in some cases other genetic elements (e.g promoter elements, epitope tags, and reporter genes) have been developed. These modules are used as PCR substrates to create products that can be targeted to specified locations in the yeast genome, thus modifying that genomic locus. We describe here a series of plasmids that contain a truncated version of the strong ADH1 promoter with and without amino-terminal 3HA and GST tags. Because these plasmids contain the same vector sequences as the GAL1 promoter plasmids, a constitutive and an inducible promoter can now be integrated with a minimal number of primers.
DeMarini
D J
DJ
Department of Comparative Genetics, SmithKline Beecham Pharmaceuticals, King of Prussia, PA 19406, USA. Douglas_J_Demarini@sbphrd.com
Carlin
E M
EM
Livi
G P
GP
D015870:Gene Expression; D005800:Genes, Fungal; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*