Designer deletion and prototrophic strains derived from Saccharomyces cerevisiae strain W303-1a.
We report the construction of Saccharomyces cerevisiae strains isogenic to W303-1a that are designed to allow efficient genetic analysis. To facilitate the generation of null alleles of target genes by PCR-mediated gene disruption, we constructed designer deletion alleles of the ARG4, TRP1 and URA3 genes. In addition, a single pair of oligonucleotide primers were designed that can be used to amplify any of several marker genes for use in PCR-mediated gene disruption. A new version of the 'reusable' hisG-URA3-hisG cassette was constructed for use in PCR-mediated gene disruption. Finally, to facilitate the formation of isogenic diploids by selection, we constructed strains that contain combinations of wild-type alleles of ADE2, HIS3, LEU2, TRP1 and URA3.
Replogle
K
K
Department of Cell and Structural Biology, University of Illinois, Urbana-Champaign, Urbana, IL 61801, USA.
Hovland
L
L
Rivier
D H
DH
D019747:Aldose-Ketose Isomerases; D000483:Alleles; D001123:Argininosuccinate Lyase; D002262:Carboxy-Lyases / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D004271:DNA, Fungal / Q000737:chemistry*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics; D006836:Hydro-Lyases / Q000737:chemistry / Q000235:genetics; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins
D017931:DNA Primers; D004271:DNA, Fungal; D005656:Fungal Proteins; D011506:Proteins; D029701:Saccharomyces cerevisiae Proteins; D002262:Carboxy-Lyases; C022378:phosphoribosylaminoimidazole carboxylase; D006836:Hydro-Lyases; C021434:imidazoleglycerolphosphate dehydratase; C087900:Arg4 protein, S cerevisiae; D001123:Argininosuccinate Lyase; D019747:Aldose-Ketose Isomerases; C490353:TRP1 protein, S cerevisiae