Saccharomyces cerevisiae strain cg1945

The bait plasmid pAS2-FHL1 and a human mammary two-hybrid cDNA library (Clontech) were sequentially transformed into Saccharomyces cerevisiae strain CG1945 according to the manufacturer's instructions (Clontech). Transformants were grown on synthetic medium lacking tryptophan, leucine and histidine but containing 1 mM 3-aminotriazole. The candidate clones were rescued from the yeast cells and re-transformed back to the same yeast strain to confirm the interaction between the candidates and the bait. The specificity of the interaction was determined by comparing the interactions between the candidates and various bait constructs. The unrelated bait plasmid pAS2-lamin C was used as a negative control.

The bait plasmid pAS2-LEF1 and a human mammary two-hybrid cDNA library (Clontech) were sequentially transformed into Saccharomyces cerevisiae strain CG1945 according to the manufacturer’s instructions (Clontech). Transformants were grown on synthetic medium lacking tryptophan, leucine, and histidine but containing 1 mM 3-aminotriazole. Approximately one million transformants were screened. The candidate clones were rescued from yeast cells and re-transformed into the same yeast strain to verify the interaction between the candidates and bait. The specificity of the interaction was examined by comparing the interactions between the candidates and various bait constructs. The unrelated bait plasmid pAS2-lamin C was used as a negative control.