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Vaccines and Antiviral Agents

High-Frequency Phenotypic Reversion and Pathogenicity of an Acyclovir-Resistant Herpes Simplex Virus Mutant

Anthony Griffiths, Donald M. Coen
Anthony Griffiths
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115
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Donald M. Coen
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115
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  • For correspondence: Don_Coen@hms.harvard.edu
DOI: 10.1128/JVI.77.3.2282-2286.2003
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  • FIG. 1.
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    FIG. 1.

    Construction of recombinant virus. Below the top two lines, which represent the tkLTRZ1 genome and the location of the tk gene (UL23), is a schematic diagram of the functional domains of TK and the location of the G string. The bottom four lines show the wild-type nucleotide sequence and a three-frame translation of this sequence, with the wild-type frame marked as the 0 frame.

  • FIG. 2.
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    FIG. 2.

    (A) Semiquantitative plaque autoradiography of viruses. The top line of values shows the approximate amounts of active TK expressed by each mutant as a percentage of that expressed by the wild-type strain KOS. The next line shows the average amount of radioactivity measured per plaque as a percentage relative to that measured for KOS. The next line presents the images of the plates. The relative level of TK activity associated with TKG7+2G is shown above the image of the plate used to ascertain this value. Relative TK activity levels were estimated from a graph (data not shown) of the relative percentage of TK polypeptide plotted against the relative percentage of TK activity in situ for LS-111/-101//-56/-46, 615.9, LS-29/-18, and tkLTRZ1 (y = 0.034x + 0.2684; R2 = 0.91). The arrowhead on the TKG7+2G autoradiograph indicates a plaque with TK+ activity. (B) Enlarged image of a single plaque that appears to have a mixed phenotype. The white arrowhead indicates a cell with a low level of TK activity, and the black arrowhead indicates a cell with a high level of TK activity.

  • FIG. 3.
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    FIG. 3.

    Plaque autoradiography of viruses isolated during the acute phase of infection with TKG7+2G. The percentage of plaques with high levels of TK activity among those with low levels of TK activity is noted. In each image, one example each of a plaque with a high level of TK activity and a TKL plaque is indicated with a black and a white arrowhead, respectively.

  • FIG. 4.
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    FIG. 4.

    Plaque autoradiography of viruses isolated following reactivation of TKG7+2G. In the image of the mixed population, one example each of a plaque with a high level of TK activity and a TKL plaque is indicated with a black and a white arrowhead, respectively.

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  • TABLE 1.

    Virus titers in eye swabs and ganglia during acute infections of mice

    VirusTiter (log meana ± SE) at indicated location and time p.i.b
    EyeTrigeminal ganglia
    Day 1Day 2Day 3Day 3
    KOS4.3 ± 0 (3)5.1 ± 0.2 (4)4.3 ± 0.3 (4)5.3 ± 0.3 (3)
    tkLTRZ15.4 ± 0.2 (3)4.0 ± 0.3 (4)2.6 ± 0.2 (4)0.3 ± 0 (4)
    TKG7+2G.15.6 ± 0.5 (2)5.5 ± 0.2 (2)2.3 ± 0.2 (2)3.9 ± 0.2 (2)
    TKG7+2G.24.6 (1)4.4 ± 0 (2)3.5 ± 0.3 (2)3.8 ± 0.3 (2)
    • ↵ a Calculated by averaging the logs of the titers.

    • ↵ b The number of samples titrated for each group is shown in parentheses.

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High-Frequency Phenotypic Reversion and Pathogenicity of an Acyclovir-Resistant Herpes Simplex Virus Mutant
Anthony Griffiths, Donald M. Coen
Journal of Virology Feb 2003, 77 (3) 2282-2286; DOI: 10.1128/JVI.77.3.2282-2286.2003

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High-Frequency Phenotypic Reversion and Pathogenicity of an Acyclovir-Resistant Herpes Simplex Virus Mutant
Anthony Griffiths, Donald M. Coen
Journal of Virology Feb 2003, 77 (3) 2282-2286; DOI: 10.1128/JVI.77.3.2282-2286.2003
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  • Top
  • Article
    • ABSTRACT
    • Construction of recombinant virus TKG7+2G.
    • Heterogeneity of TK activity of virus carrying a double-G insertion.
    • Enrichment of virus with high levels of TK activity during acute replication in the mouse.
    • Reactivation from latency.
    • What accounts for the low level of TK activity of most TKG7+2G plaques?
    • What causes the high frequency of phenotypic reversion?
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

Acyclovir
antiviral agents
Simplexvirus
thymidine kinase

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