This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Flock House Virus (FHV) is a simple, non enveloped virus encoding two gene products from a bipartite (RNA 1, -3000 nucleotides encoding the polymerase and RNA 2 -1500 nucleotides encoding the capsid protein) ssRNA genome. We seek to follow virus particle entry into Drosophila Line I (DLI) cells with fluorescently labeled particles employing fluorescence microscopy to localize particles during (anticipated) endocytosis. Employing fluorescently labeled antibody to the coat protein we seek to identify the locations of coat protein synthesis and virus assembly within the cell. Employing fluorescently labeled antibody to the polymerase and cDNA oligomers (with fluorescent tags) to RNAI and RNA2 we seek to identify locations of RNA synthesis within the cell. We also seek to study viral cytopathic effects on DL1 cells by a time course study of viral infection using negative stain electron microscopy and computer tomography.
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