Criollo Fajardo, Melisa M.

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    Determining the role of FAP1 in S. cerevisiae, cultivated in nitrogen-limited media
    (2024-07-09) Criollo Fajardo, Melisa M.; Parés Matos, Elsie; College of Arts and Sciences - Sciences; De Jesús Ruiz, Marco; Román Velázquez, Félix; Department of Chemistry; Perez Muñoz, Fernando
    FAP1 is a cytoplasmic protein found in Saccharomyces cerevisiae. During Rapamycin treatment, FAP1 binds Fpr1p for conferring resistance to yeast cells. Rapamycin is lethal to yeast cells because it can also bind Fpr1p, and this complex interferes negatively with the TORC1 function, which is responsible for nutrient-regulated gene expression. Currently, FAP1 function remains poorly understood. It has been suggested that under nutrient-deficient conditions, the fap1 gene is over-expressed and becomes essential for yeast to survive. This work identified the expression levels of the fap1 gene when Saccharomyces cerevisiae is exposed in Nitrogen-limited media (NLM) to 0 h, 3 h, 6 h and 9 h. After harvesting yeast cells, total RNA was extracted and then treated with Reverse Transcriptase to create complementary DNA (cDNA), yielded cDNA with concentrations ranging from 1939 ng/µL (0 h), 1608 ng/µL (3 h), 1845 ng/µL (6 h) and 1521 ng/µL (9 h). Subsequently, qPCR experiments were performed using cDNA samples from yeast cells grown in YPD (control) at 0 h. This served to validate the amplification efficiency of the fap1 gene relative to the endogenous fba1 gene, establishing the necessary conditions for acquiring experimental cycle threshold (CT) values. fap1 cDNA exhibited an increased expression in each of the three experimental media studied here (NLM) as time progresses.