Calibration and illustrative results from the pool competition approach. (a) Pool competition results for selection for canavanine resistance. The abscissa indicates the proportion of each strain in the untreated pool. When treated (y-axis) the ability of the can1Δ/can1Δ diploid to resist canavanine confers a major growth advantage to the strain such that it outcompetes all others to become the most abundant strain. Pink and green lines (± 1000 y-translation of parity line) detail the boundary within which 98% of deletants are found when comparing untreated controls, so providing a noise estimate. Indigo and cyan lines indicate treatment/control ratios of 2 and 3. Blue diamonds denote all deletants and black stars identify strains deleted for transporter genes. AFU = mean arbitrary fluorescent units measured for the TAG4 arrays of treated (canavanine) and untreated (control) competitions between the pools of homozygous deletants. (b) Growth curves of wild type (ydl227cΔ/ydl227cΔ) yeast strains in the presence of 10 μM of canavanine and increasing concentration of the competitor arginine. (c) Comparison of the maximum specific growth rate achieved by the WT strain in the presence and absence of canavanine illustrates the cytotoxic effect of the drug. The protective effects of various concentrations of arginine, one of the native substrates of Can1p, are shown over the drug concentration range (0 mM to 100 mM). A similar growth rate advantage to deleting CAN1 is obtained by adding 250 μM of arginine. Error bars = standard error of the mean; n = 3.
Lanthaler et al. BMC Biology 2011 9:70 doi:10.1186/1741-7007-9-70