Construction of recombinant herpes simplex virus type I expressing green fluorescent protein without loss of any viral genes.
For use in various applications in research on herpes simplex virus type 1, we attempted to generate recombinant HSV-1 expressing green fluorescent protein (GFP) without any loss of viral genes. Our results were as follows. (i) A recombinant HSV-1 (YK333), in which a GFP expression cassette driven by the Egr-1 promoter was inserted into the intergenic region between UL3 and UL4, was constructed. (ii) YK333 replicated as well as wild-type HSV-1 F strain in Vero cells. (iii) As one application of the recombinant YK333 for research on HSV-1, we developed a system to detect anti-herpetic activity, termed a fluorescence-based anti-viral assay. The 50% inhibitory concentration of ganciclovir for YK333 determined using our newly developed assay was comparable to that determined using a plaque reduction assay. YK333 will be a convenient tool for herpes simplex virus research, including such applications as monitoring of viral replication in vitro and in vivo, and rapid screening of potential anti-herpetic agents.
Tanaka
Michiko
M
Department of Pathology, National Institute of Infectious Disease, 1-23-1 Toyama, Sinjyuku-ku, Tokyo 162-8640, Japan.
Kodaira
Hiroshi
H
Nishiyama
Yukihiro
Y
Sata
Tetsutaro
T
Kawaguchi
Yasushi
Y
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D021901:DNA, Intergenic; D005453:Fluorescence; D015774:Ganciclovir / Q000494:pharmacology; D015967:Gene Expression Regulation, Viral; D049452:Green Fluorescent Proteins; D018259:Herpesvirus 1, Human / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014162:Transfection; D014709:Vero Cells; D010948:Viral Plaque Assay; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D021901:DNA, Intergenic; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D015774:Ganciclovir