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.
Boldogköi
Z
Z
Laboratory of Molecular Virology, Agricultural Biotechnology Center, PO Box 411, H-2101 Gödöllö, Hungary.
Erdélyi
F
F
Sik
A
A
Freund
T F
TF
Fodor
I
I
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