Date of Award

2011

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biology

First Advisor

Foushee, Doretha B. Dr.

Abstract

Infectious pancreatic necrosis virus (IPNV) is a bi-segmented, double stranded RNA virus that causes high mortality in young salmonid fish worldwide. As an obligate intracellular parasite, the first stage of the IPNV infectious cycle is the binding of the virus to a receptor in the host cell membrane. Recently, research has focused on identifying the IPNV receptor, and several scientists have reported binding of IPNV to a protein of approximately 220-250 kDa. The focus of this research was to investigate and reaffirm the identity of the IPNV receptor(s) for IPNV serotypes VR-299 and Sp in Chinook Salmon Embryo (CHSE-214) cells and Rainbow Trout Gonad (RTG-2) cells. We hypothesized that both serotypes VR-299 and Sp utilize the same receptors. Membrane proteins from CHSE-214 and RTG-2 cells were extracted using the Calbiochem Proteo Extract Native Membrane Protein Extraction Kit. The Virus Overlay Protein Binding Assay (VOPBA) was used to identify IPNV-protein interactions. Our results confirmed the binding of IPNV serotypes Sp and VR-299 to a protein approximately 250kDa in the CHSE-214 and RTG-2 cells. However, our results also revealed binding of both IPNV serotypes Sp and VR-299 to two additional proteins of approximately 60kDa and 65kDa in the CHSE-214 and RTG-2 cells and a third protein at 140kDa in CHSE-214 cells.

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