Stephanie Nguyen
"I applied to USP at the encouragement of my research advisor with the aim of being able to continue my research during the summer. During the academic year, my goal is to hopefully complete my research in time to submit for publication."
Major
Microbiology
Minor
Business Administration
Research Interests
Microbiology.
Academic and Other Awards
- University Scholars Program Scholarship (2011-2012)
- Bob Graham Civic Scholar
Organizations
- UF Campus Kitchen
- Club Alligator Swim Team
Volunteer
Making meals with Campus Kitchens!
Hobbies and Interests
Research Description
G-Quadraplex DNA Structure Effect on Transcription-Coupled DNA Repair
DNA sequences that can form structures that deviate from the canonical, helical duplex-B DNA structure often correspond to genomic regions that are susceptible to genetic instability, an early contributing factor in human cancer development. DNA repair processes are necessary components in the mutagenic process occurring at these non-canonical structures, and transcription through these regions promotes these structures’ formation. Because of the unusual structural features, these regions may be recognized as form of DNA damage by nucleotide excision repair (NER). Transcription-coupled DNA repair, a subpathway of NER, initiates as a result of the transcription arrest at the DNA structure sites and facilitate recruitment of repair proteins. To test this model, the effects of transcription and excision repair will be investigated in sequences that form quadruplex DNA structures identified in the c-myc gene. G-quadruplex (G4) DNA forms in DNA sequences with guanine repeats and consists of planar arrays of hydrogen-bonded guanines. The G4 forming sequence from the c-myc promoter is cloned downstream of the T7 or the Adenovirus major late promoter for our studies. A digest with restriction enzyme BsgI will confirm formation of the desired structure. BsgI will not cut the plasmid if the G4 structure is present, since the cut site is within the structure. Next, transcription is run on the plasmid using T7 RNA polymerase, a single subunit enzyme, to measure the extent of the transcription arrest. T7 RNA polymerase is a viral polymerase that is easier to use than RNA polymerase II. After transcription arrest is confirmed with the T7 RNA polymerase, transcription is repeated with RNA Polymerase II. HeLa whole cell extracts are then added to introduce repair factors to the system, and repair assays are run to measure the efficiency and fidelity of the repair process.