Construction of a recombinant herpesvirus expressing the jellyfish green fluorescent protein

Here we report the insertion of a synthetic version of the cDNA encoding the jellyfish (Aequorea victoria) green fluorescent protein (gfp(h)) into the genome of pseudorabies (Aujeszky's disease) virus (PrV). A putative latency promoter (P-LAT) located at the inverted repeat region of the PrV genome was chosen as the target site for the insertion. Recombinant viral DNA designated as vLAT-gfp was generated as a result of homologous recombination between the transfected viral DNA and a plasmid containing the GFP-expression cassette flanked by viral sequences homologous to the target region. Plaques containing recombinant virus were selected visually using a fluorescent microscope. We demonstrated a GFP-expression in infected neurons of rat brain which showed normal morphology at early stage of viral infection by monitoring fluorescent light emission. Copyright (C) 1999 John Wiley & Sons, Ltd.