Figure 1
From: Removing auto-activators from yeast-two-hybrid assays by conditional negative selection
Synthesized pGAL2-URA3 plasmid and outline of the method. (A) pRS305k-pGAL2-URA3 plasmid harboring URA3 driven by pGAL2 promoter. (B) Scenario where the yeast cell does not contain an auto-activator (AA). URA3 and HIS3 are not expressed, and FOA is not converted into FU. (C) Scenario where an AA activates the expression of URA3 and HIS3, resulting in the conversion of FOA into toxic FU by URA3. (D) Description of the AA-removal method, where a starter culture containing AAs is treated by FOA. Since FOA can inhibit or kill yeast, the growth of AAs should be severely delayed or outright stopped by FOA.
