FOREIGN PROTEIN TAGGED WITH C-TERMINAL PIV5 NP PROTEIN AND NUCLEAR LOCALIZATION SEQUENCE ALLOWS FOR PACKAGING OF CARGO INTO PIV5 VIRUS-LIKE PARTICLES: AN EXPLORATION OF THE M-NP INTERACTION AS A STEP FORWARD IN TECHNOLOGY FOR PRODUCTION OF CARGO-LOADED VLPS
Open Access
Author:
Lorenz, Fred Jeffrey
Area of Honors:
Veterinary and Biomedical Sciences
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
Anthony Paul Schmitt, Thesis Supervisor Dr. Kumble Sandeep Prabhu, Thesis Honors Advisor
Keywords:
Virus VLP Mumps Measles Paramyxovirus Virus-like particles PIV-5 virology Veterinary and Biomedical Sciences Pathobiology Infectious disease microscopy western blot
Abstract:
Paramyxoviruses are negative sense single stranded RNA viruses associated with many potentially fatal infections of the respiratory tract. Although paramyxoviruses pose alarming medical, agricultural, and hence economic burdens, a very promising area of research poses to utilize their viral machinery to our advantage in the delivery of proteins of interest to cells by way of virus-like particles (VLPs). To date, our lab has discovered that the C-terminal end of paramyxovirus nucleocapsid protein (NP) is critical for VLP production. In addition, fusing a foreign protein with the sequence derived from the NP C-terminal end allowed it to be packaged into budding VLPs. This thesis builds upon previous research by confirming that the addition of a nuclear localization sequence to foreign protein (here GFP.NLS) alters its localization to the nucleus of cells, and demonstrating that GFP.NLS fused with the 20-residue sequence derived from the viral NP C-terminal end (GFP.NLS.NP20) can still be incorporated into VLPs despite its increased confinement to the nucleus. However, expression of M did not significantly alter the amount of nuclear GFP.NLS.NP20, which presents a disconnect between the mechanism of delivery to the nucleus and subsequent propensity to bud.
