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 (gfph ) into the genome of pseudorabies (Aujeszky's disease) virus (PrV). A putative latency promoter (PLAT) 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.
Laboratory of Molecular Virology, Agricultural Biotechnology Center, PO Box 411, H-2101 Gödöllö, Hungary.
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Boldogköi
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Erdélyi
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Sik
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Freund
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I
Fodor
D000818:Animals; D001921:Brain / Q000166:cytology*; D002460:Cell Line; D018076:DNA, Complementary; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D011558:Herpesvirus 1, Suid / Q000235:genetics* / Q000302:isolation & purification; D008164:Luminescent Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D009474:Neurons / Q000166:cytology*; D011401:Promoter Regions, Genetic; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D011995:Recombination, Genetic; D037401:Scyphozoa; D013552:Swine; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins