This video demonstrates the generation of influenza virus-like particles (VLPs) in a mammalian system. Upon expressing three eukaryotic expression vectors — encoding different viral structural proteins — in suitable mammalian host cells, the expressed viral proteins self-assemble into VLPs on the host plasma membrane and release via budding from the cell surface.
Protocol
1. Mammalian Expression System for Generation of Influenza H3N2 VLP Subclone viral structural glycoproteins hemagglutinin (HA), neuraminidase (NA), and human immunodeficiency virus (HIV) Gag (group-specific antigen) p55 into eukaryotic expression vectors, such as previously described pTR600. Amplify DNA in chemically competent Escherichia coli (e.g., DH5α) and isolate transfection-grade plasmid using a plasmid purification kit per manufacturer's instructio…