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. 2018 Jul 30;8(5):772–783. doi: 10.1016/j.apsb.2018.07.006

Figure 1.

Figure 1

Detection of L10–L12 interaction using yeast two-hybrid assay. (A) The rationale for high throughput screen using yeast two-hybrid system. L10–L12 interaction reconstitutes the function of the Gal4 protein and enables the expression of reporter genes, ADE2, HIS3, and LacZ. The disruption L12–L10 interaction by a compound inhibits yeast cell growth in SD/Ade/His dropout medium and β-gal will not be produced. (B) The growth and β-gal activity of yeast cells expressing various combinations of BD and AD fusions. The combination of the different plasmids in AH109 strains is shown on the right. Among them, strains 1 and 6 are the positive and negative controls, respectively. The left panel shows the growth of yeast cells with indicated plasmids on a SD/Leu/Trp/Ade/His dropout plate. The right panel shows the β-gal activity of indicated strains. (C) Quantification of β-gal activity in yeast cells containing various combinations of plasmids. The results are the average from triplicate assays. (D) The expression of L12 and L10 proteins in yeast cells. L12 and L10 fusion proteins were expressed in yeast cells, and the expression was detected using anti-HA and anti-Myc antibodies.