Figure 2.

Graphic representation of the reporter genes and plasmids used in the two-hybrid screen shown in Figure 1B. The screen reported here includes two reporters and two plasmids. (A) A reconstituted transcription factor will activate transcription of the HIS3 gene and simultaneously activate transcription of the lacZ gene. The HIS3 reporter can be selected for growth on plates containing 3-amino-triazole (3AT), and the lacZ reporter can be assayed by performing a ß-galactosidase assay that will cause blue color formation in yeast. (B) The plasmids transformed into the yeast for the two-hybrid screen. The Galcdk6 plasmid encodes the Gal4 DBD fused to cdk6. This plasmid also encodes the LEU2 gene for use in auxotrophic selection. The second plasmid encodes the cDNA library fused to a sequence encoding a TAD. This plasmid also encodes the marker TRP1, a gene involved in tryptophan biosynthesis.