Quantitative Analysis of Gene Expression

Gene expression analysis was performed according to previously described method (Demuyser et al., 2017 (link)). The cells were grown to mid-exponential phase in SC medium supplemented with the carbon source of interest. Cells were washed with ice-cold Milli-Q water, frozen in liquid nitrogen and kept at −80°C. The cell pellet was dissolved in TRIzol (Thermo Fisher), and cells were lysed by fast prepping with glass beads. Afterward, RNA was isolated using chloroform, isopropanol, and 70% ethanol. The RNA was treated with DNase enzyme (New England Biolabs), to remove present DNA fractions, and converted into cDNA by using the iScript cDNA synthesis kit (Bio-Rad). To perform the quantitative PCR, a Go-Taq polymerase (Promega) and a StepOnePlus machine (Thermo Fisher) were used. The results were analyzed by making use of the qBasePlus software (Biogazelle).