Date of Award
Winter 3-22-2024
Degree Type
Thesis
Degree Name
Master of Science (MS)
Department
Biological Science
First Advisor
Sarah Connolly, PhD
Second Advisor
Margaret Bell, PhD
Third Advisor
John Dean, PhD
Abstract
A sequence of protein-protein interactions between four glycoproteins (gB, gD, gH, and gL) are sufficient and necessary for entry of herpes simplex virus (HSV) into a host cell. The mechanism for entry is conserved across all herpesviruses, specifically gH/gL and gB (Connolly et al., 2011), therefore studying HSV-1 entry can give insight into other herpesviruses. Co-immunoprecipitation (coIP) is an appealing technique for studying protein interactions required for entry in HSV-1. coIP is a promising method because wild-type protein complexes remain intact, and cells can be transfected with multiple combinations of proteins for isolation. coIP can be challenging because the interactions observed may be low affinity and/or transient. To establish optimal conditions for coIP, proteins were first analyzed for expression using western blotting. Secreted HSV-1 proteins were determined by western blot analysis and a coIP protocol was optimized. Using secreted proteins is an advantage because a detergent is not needed, and detergent can disrupt protein-protein interactions. The findings show that a coIP did not demonstrate a protein-protein interaction for HSV-1.
Recommended Citation
Consalvo, Angelina, "Assay Optimization to Capture Herpesvirus Entry Protein Interactions" (2024). College of Science and Health Theses and Dissertations. 612.
https://via.library.depaul.edu/csh_etd/612
SLP Collection
no