Deletion of Oxidative Stress Genes in C. glabrata

The following genes were subjected to deletion, catalase 1 (CAT1, CAGL0K10868g), glutathione oxidoreductase (GRX2, CAGL0K05813g), manganese superoxide dismutase (SOD2, CAGL0E04356g), and three transcription factors playing role in oxidative stress responses, namely SKN7 (CAGL0F09097g), MSN4 (CAGL0M13189g), and YAP1 (CAGL0H04631g), and ICL1 (CAGL0L09273g). Knock-out mutants were created in house using a previously described protocol^{65 (link)}, in which the open reading frame of the gene of interest was replaced by nourseothricin (NAT) resistance cassette. The knock-out construct was generated by using Ultramer primers (∼80–100 bps) containing homology regions with NAT and with regions flanking GOIs. Competent cells were created by log-phase grown C. glabrata cells using Frozen-EZ Yeast Transformation Kit (Zymo Research) and transformation followed an electroporation-based protocol described previously^{67 (link)}. The colonies growing on YPD plates containing NAT were subjected to PCR and sequencing using diagnostic primers listed in Supplementary Table 3 (the primers used in the current study were manufactured by Integrated DNA Technologies and Sanger Sequencing carried out by Genewiz). Two independent knock-out mutants were used for each experiment.