Disintegrator vectors for single-copy yeast chromosomal integration.
Vectors were developed for two-step chromosomal integration of reporter genes or expression constructs. With these vectors, integration produces a disruption of the ADE8, LYS2, MET15, LEU2, HIS3 or FCY1 genes, and integrants can be easily identified by replica-plating on selective media. Integration using these 'disintegrator' vectors produces a single-copy integration of the construct of interest at the junction of the marker deletion, and removes the additional plasmid sequences. Importantly, the integrated constructs do not contain flanking sequence duplications, and therefore should be highly stable. Each of the vectors was shown to reliably integrate a TEF1-KAN expression cassette and/or GAL1-HIS3 and STE12-LacZ reporter genes.
Sadowski
Ivan
I
Department of Biochemistry and Molecular Biology, Molecular Epigenetics, Life Sciences Centre, University of British Columbia, Vancouver, BC, Canada. sadowski@interchange.ubc.ca
Su
Ting-Cheng
TC
Parent
Jennifer
J
D015825:Chromosomes, Fungal; D017930:Genes, Reporter; D005822:Genetic Vectors; D016254:Mutagenesis, Insertional / Q000379:methods*; D010957:Plasmids / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D014170:Transformation, Genetic