Finding genes that resist thioconazole from new generation triazole group antifungal drugs

Finding Genes That Resist Thioconazole From New Generation Triazole Group Antifungal Drugs Aim: New developments in medicine, like bone marrow transplantation, chemotherapy, and liver and kidney transplantations, cause an increase in the number of patients requiring intensive care due to the broad-spectrum antibiotics and corticosteroids used in these treatments, prolonged intensive care processes, and the morbidity and mortality of systemic fungal infections due to the immunosuppressants used to prevent organ rejection in organ and tissue transplants. Antifungal susceptibility tests take a long time to perform, which causes delays in giving the necessary treatment to patients and promotes the ineffective use of antifungals. Our aim is to determine how the cells become resistant to thioclosole (TIO), which is used in the treatment of fungal infections, by using genetic/genomic methods that can be applied in yeast. Material and Method: In our study, we determined the MIC dose for TIO in YPD agar medium for the BY4741 S. cerevisiae yeast strain. For the purpose of screening, we spread gDNA-transformed cells on YNB plates that contained lethal doses of the drug and isolated plasmids from the isolates that were less sensitive to the drug. We used it to transform fresh wild-types to prove that they were responsible for the growth in the presence of the drug. After confirmation, the plasmids were sequenced using the Yep13F1 primer. Results: The resistant colony was found to contain a DNA region on the 12th Chromosome of Maya. This DNA sequence was used to perform a BLAST analysis in the Yeast Genome Database (https://www.yeastgenome.org) and it was found to correspond to the gene named YLR117w. Conclusion: We observed that overexpression of the YLR177w gene reduced TIO sensitivity. We analyzed the deletion mutant without YLR177w in terms of its ability to grow in the presence of the drug, but we observed that cells were not susceptible to TIO in the absence of YLR177w.