Discipline: Chemistry and Chemical Sciences
Room: Park Tower 8217
Ulyssa Robertson - Central Piedmont Community College
Co-Author(s): Katelyn Erickson, University of North Carolina at Charlotte; Jerry Troutman, University of North Carolina at Charlotte
Bacterial polysaccharides play specific roles in the protection and virulence of bacteria. Therefore polysaccharides serve as possible target molecules for sensors, or other therapeutics. Polysaccharides could be general to a bacteria family but also specific within that family due to the surface glycan structures. The goal of this project is to develop a tool for a more specific therapeutic treatment via a directed approach. The targeting approach stems from the use of a DNA aptamer that like an antibody, is specific for a biomolecule. Glycan specific aptamers will be selected based on its binding activity to the surface of enterobacteriaceae family of Escherichia coli, specifically for the ECA polymer by selecting against a strain that does not contain the ECA polymer. A library of 10^12 oligonucleotides was obtained and aptamer selection progressed through systematic evolution of ligands by exponential enrichment (SELEX). The pools were amplified by the polymerase chain reaction (PCR), a process to help make several copies was performed after each selection, and viewed using gel electrophoresis. Through the course of the 10- weeks I completed 5 selections which was the halfway point of decreasing my DNA pool. I progressed with the DNA pool and TOPO cloning was performed for fast and efficient analysis of PCR- amplified DNA. In future research, I plan to do more selections to further enrich the DNA pool to bind to ECA sugar polymer, and use a fluorescent forward primer to detect aptamers binding to my target cell.
Funder Acknowledgement(s): National Science Foundation; NanoSURE REU program
Faculty Advisor: Jerry Troutman, email@example.com
Role: I conducted the research with guidance from a graduate student (Katelyn Erickson) and faculty mentor (Jerry Troutman)