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Identification of mutants with altered PHO84 expression in low and high Pi conditions.

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posted on 2017-05-17, 17:30 authored by Joonhyuk Choi, Abbhirami Rajagopal, Yi-Fan Xu, Joshua D. Rabinowitz, Erin K. O’Shea

(A) Generation of single mutants harboring the PHO84 reporter with the SGA method. The PHO84 reporter consists of PHO84 promoter-driven Venus and TEF2 promoter-driven mCherry. Each single mutant in the library denoted by xxxΔ is kanamycin (G418)-resistant. (B) The distributions of the PHO84 reporter levels in single cells in the pho81Δ and pho80Δ strains. Log2 intensity ratio of Venus to mCherry (log2(YFP/RFP)) was used to quantify the PHO84 expression level. (C, D) The PHO84 reporter levels of single mutants in the library measured in 50 μM Pi and 1 mM Pi conditions. The PHO84 reporter level of each mutant was normalized to that of the wild type value in each Pi concentration (Materials and methods). Red dashed lines in (C) and (D) indicate the PHO84 reporter levels with p-values less than 0.001 estimating the maximum range of the PHO84 reporter levels that the wild type exhibits in each Pi concentration. The mutants in black are previously identified mutants and the one in red is identified in this study.