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Fig. 1 | BMC Biology

Fig. 1

From: A yeast two-hybrid system for the screening and characterization of small-molecule inhibitors of protein–protein interactions identifies a novel putative Mdm2-binding site in p53

Fig. 1

p53 interacts with Mdm2 in the yeast two-hybrid (Y2H) assay. a Schematic showing the use of the Y2H assay in identifying interacting proteins (left panel) and inhibitors of protein–protein interactions (right panel). b Log-phase cultures of AH109 yeast cells containing plasmids encoding either Gal4 AD-p53/Gal4 BD-Mdm2, Gal4 AD-p53/Gal4 BD, Gal4 AD/Gal4 BD-Mdm2, or Gal4 AD/Gal4 BD were washed in water and plated at different dilutions on non-selective (-Leu-Trp) and selective (-Leu-Trp-Ade-His) plates and incubated at 30 °C for 3 days. c Overnight cultures of AH109 yeast cells containing plasmids encoding either Gal4 AD-p53/Gal4 BD-Mdm2, Gal4 AD-p53/Gal4 BD, Gal4 AD/Gal4 BD-Mdm2, Gal4 AD/Gal4 BD, Gal4 AD-p53-F19A/Gal4 BD-Mdm2, or Gal4 AD-p53(Δ42)/Gal4 BD-Mdm2 in non-selective medium were washed in water and inoculated into selective and non-selective medium at OD600 = 0.2 in duplicates. For each strain, growth as measured by average OD600 of duplicate cultures is plotted against time. Ends of the vertical bar indicate the OD600 values of the duplicate cultures

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