TY - JOUR TI - Creation and characterization of VZV recombinant strain ORF7Δ in a novel RFP-ORF23 fusion BAC construct DO - https://doi.org/doi:10.7282/T3JQ0ZBC PY - 2014 AB - The Alphaherpesvirus varicella zoster virus (VZV), while widely vaccinated for in the United States, still remains an important human pathogen. The bacterial artificial chromosome method (BAC) was used to make a deletion mutant of the ORF7 protein, implicated in the observed tissue tropism exhibited by VZV, in an RFP-ORF23 fusion BAC. For this experiment, a novel double-selection cassette using galactokinase and zeocin as markers was developed to attempt to circumvent the shortcomings of single selection additions to a BAC. Red fluorescent protein coupled to ORF-23, a viral tegument protein, was used to confirm positive infection of ARPE cells, and should allow direct visualization of the viral life cycle with fluorescence microscopy illuminating where in VZV’s life cycle ORF7 is required for viral release. The virus exhibited a slow-growing phenotype typical of ORF7 deletion mutants in relation to wild-type. Further research will aim to produce a suitable vaccine strain called ORF7S, and visualize this clone as it moves through differentiated neuronal cells. KW - Microbiology and Molecular Genetics KW - Varicella-zoster virus KW - Bacterial artificial chromosomes KW - Recombinant baculoviruses LA - eng ER -