Comprehensive Yeast Strain and Plasmid Protocols

A list of Saccharomyces cerevisiae strains and plasmids is provided in S4 Table. For strain maintenance and construction, strains were grown at 30°C under standard conditions. ncRNA single deletion strains used this study were taken from the ncRNA deletion collection created by Parker et al [10 (link), 78 (link)]. Deletion mutants were maintained on Yeast extract Peptone Dextrose Agar (YPDA) containing 200 μg/mL G418. Double deletion mutant strains were constructed by substituting the candidate SUT locus with the natNT2 cassette and were maintained on YPDA containing 100 μg/mL clonNAT.
For construction of strains ectopically expressing particular SUTs, isogenic wild-type and ncRNA deletion mutant strains cells were transformed with pRS416-Gal1-Cyc1 overexpression plasmid containing the ncRNA of interest. Resulting strains were maintained in a synthetic minimal media lacking uracil (SD-Ura: 1X Yeast Nitrogen Base (YNB) (Formedium); 1X Complete Supplement Mixture (CSM)–Ura (Formedium); 2% (w/v) glucose). For phenotypic rescue studies, strains were grown to an optical density at 600 nm (OD₆₀₀) of 0.5 in YP (1% yeast extract, 2% peptone) medium supplemented with 2% raffinose (YPRaf) at 30°C and induced with YP medium containing 2% galactose (YPGal) for 2 hours before being harvested for spot test assays.