Host-Viral Protein-Protein Interactions in Influenza virus Infections
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Influenza virus infection remains a worldwide problem today. Pathogenicity can be attributed in part to the changing protein structure of the virus, allowing it to evade the host immune system. For example, the hemagglutinin (HA) and neuraminidase (NA) proteins used in the subtyping of influenza strains; their roles and interactions with host proteins having been extensively studied. Studying host-viral protein-protein interactions is important in understanding the virus replication process, and still there are many other key proteins worth investigating to further our knowledge in influenza research and potentially target new treatment. This research project will focus on the influenza virus protein NS1, a non-structural protein involved in the viral replication cycle, exploring host-viral protein-protein interactions throughout the replication process in an attempt to characterize the exact proteins NS1 requires during infection. The student will primarily be using a non-pathogenic influenza lab strain A/PR/8/34 (H1N1 subtype) with a mammalian cell line of the Madin-Darby Canine Kidney (MDCK) epithelial type. The objective of the student will be to learn proper and aseptic cell culturing technique, methods for protein analysis such as co-immunoprecipitation to “pull-down” proteins of interest using monoclonal antibodies directed against NS1, which the student will be purifying from murine lymphocytes, as well as polyacrylamide gel electrophoresis (SDS PAGE) to separate proteins, and finally western blotting. Any interacting proteins seen will then be sent for analysis using mass spectrometry to determine their exact identities for further study.