Fig. 3
SCRaMbLE of CBS5829-syn(V)X improves caffeine tolerance. a Serial dilution assay comparing the growth of SCRaMbLEd S. paradoxus CBS5829-synX (yMS637, yMS638, yMS639, yMS640, yYW184, and yYW185) or CBS5829-synVsynX (yYW186, yYW187, yYW195, and yYW196) strains to their non-SCRaMbLEd CBS5829-synX parent (yMS521) on high caffeine YPD plates. b The POL32 gene with 500 bp upstream/300 bp downstream sequence was cloned from BY4741 into the episomal plasmid pRS416 and the resulting plasmid pRS416-POL32 transformed into yMS521. These strains were compared via serial dilution assay to the SCRaMbLEd strain yYW185 transformed with pRS416, yMS521 transformed with pRS416, or BY4743 transformed with pRS416 on SC–Ura + 4 mg/mL caffeine. c yMS521 transformed with pRS416, yMS521 transformed with pRS416-POL32, yYW184 transformed with pRS416, and yYW185 transformed with pRS416 were all grown in liquid SC–Ura media overnight and then diluted to a starting A600 of 0.1 in either SC–Ura + 4 mg/mL or 5 mg/mL caffeine and cultured in a 96-well plate reader with shaking. Optical density measurements were taken every 10 min and used to calculate doubling time. Error bars shown are mean and standard deviation from four technical replicates. One-way ANOVA with multiple comparisons was used to assess difference between yMS521 + pRS416 and other samples (****p < 0.0001). Variance between the groups was determined to be similar
