Level of Education of Students Involved

Undergraduate

Faculty Sponsor

Patrice Bouyer

College

Arts and Sciences

Discipline(s)

Biology Department

Presentation Type

Poster Presentation

Symposium Date

Spring 4-25-2024

Abstract

Candida albicans is a commensal fungal pathogen, and to infect the human body, it must penetrate the intestinal mucosal barrier by changing its morphology from yeast to filamentous. The risk of candida infections increases during pregnancy and estrogen levels are high during pregnancy. Thus, we hypothesized that estrogen could act as a signal for C. albicans to become filamentous. In an ongoing study, we have looked at the effect of estrogen on filamentation, but our approach was qualitative rather than quantitative and could not clearly establish the effect of estrogen on filamentation. To get a better understanding of the effect of estrogen we have decided to use qPCR to quantify the gene expression of HWP1 involved in filamentation. In the literature, it is reported that extracting RNA from fungi is challenging because of the cell wall. In our experiments, we have compared 4 extraction methods a) snap freeze/Trizol, b) zirconium beads/Trizol, c) snap freeze/beads/Trizol, and d) hot Formamide-EDTA (FAE) extraction. Methods “a, b, and c” yield poor RNA extraction ~50ng/µl, measured with a nanodrop. Method “d” on the other hand resulted in 300 ng/µl RNA yield and a good RNA quality on agarose gel. Using the total RNA obtained from method “d”, we generated cDNA by RT-PCR and successfully PCR HWP1 in our samples. In conclusion, FAE total RNA extraction is an effective method to obtain good RNA from C. albicans and perform PCR.

Biographical Information about Author(s)

Sara- Undergraduate Valparaiso University

George- Undergraduate Ivy Tech

Patrice - associate professor

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