Neurotropic herpesvirus envelopment and microtubule-mediated transport

The proposed research requires implementation of several technical procedures that my lab has substantial experience with and performs on a regular basis. In addition, the work requires handling of herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV, a veterinary herpesvirus) using BSL2 guidelines. I have been studying these herpesviruses for over a decade and have a BSL2 facility dedicated to herpesvirus propagation. I will produce recombinant HSV-1 and PRV that encode mutations in the coding sequences of viral genes and insertions of fluorescent protein coding sequences. These viruses will be examined for propagation and cell-cell spread kinetics using plaque assays and single-step growth curves. We will also perform imaging of living infected cells (primary neurons and epithelial cells) using fluorescence microscopes in our BSL2 facility designed for this purpose. Imaging will be used for analysis of virus assembly, intracellular transport, and egress.