Dna Methylation In Histoplasma Capsulatum
Abstract Category: Science
Course / Degree: Bachelors of Science Emphasis in Molecular Biology
Institution / University: University of Southern Mississippi, United States
Published in: 2011
Histoplasma capsulatum is the causative agent of histoplasmosis, a respiratory disease prevalent in the Mississippi and Ohio River Valleys. The organism is found as a multicellular mold in soils contaminated with bird or bat excreta (at 25° C). Conversion to the unicellular parasitic yeast takes place when aerosolized spores are inhaled into the lungs (at 37° C). Hence, the mold-to-yeast transition is vital for the pathogenesis of the disease. Therefore, the molecular aspects of this phenotypic conversion are essential. In eukaryotic organisms, DNA methylation plays a crucial role in controlling gene expression. DNA methylation typically decreases gene transcription; which is essential for normal development and required for the differentiation of cell types. On going studies in other organisms state that DNA methylation plays a role in gene expression. However, there are no previous methylation studies in Histoplasma capsulatum.
The main objective of this project is to assess the role of DNA methylation in Histoplasma capsulatum. To achieve this, genomic DNA was extracted from both the yeast and the mold forms of the Downs strain. Thereafter, the genomic DNA was digested with the isoschizomers HpaII and MspI. As an independent assay of DNA methylation, Histoplasma capsulatum (Downs strain) was grown in the presence of the cytosine analogue (5-Azacytidine) in order to assess phenotypic changes resulting from DNA demethylation. Furthermore, DNA methylation was quantitatively analyzed using an enzyme linked immunosorbent assay (ELISA). The results indicate that 68.3% of the Downs yeast genomic DNA was methylated whereas only 42.2% of the Down mold genomic DNA was methylated. Future investigations should include a bisulfite DNA treatment in order to obtain basepair-resolution DNA methylation pattern.
Thesis Keywords/Search Tags:
Epigenetics, ELISA, DNA Methylation, Rupesh Patel, Wiggins, USM, UMMC, Jackson, Histoplasmosis, Dedicated to William Ross Dean, Ross Dean, Histoplasmosis
This Thesis Abstract may be cited as follows:
http://www.openthesis.org/documents/Dna-methylation-in-histoplasma-capsulatum-600951.html
Submission Details: Thesis Abstract submitted by Rupesh Patel from United States on 07-Aug-2011 10:07.
Abstract has been viewed 2890 times (since 7 Mar 2010).
Rupesh Patel Contact Details: Email: rpatel1@umc.edu
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