isolation of a lacZ insertion mutant and association of the UL5 gene product with other members of the helicase-primase complex.

isolation of a lacZ insertion mutant and association of the UL5 gene product with other members of the helicase-primase complex.

The corneal discs of 41 patients with scarring reminiscent of herpetic infection were organ cultured for HSV isolation. To evaluate the anti-HSV activity of ASP2151, susceptibility testing was performed on viruses isolated from patients participating in a placebo- and valacyclovir-controlled proof-of-concept phase II study for recurrent genital herpes. When both methods were used, HSV was identified in 97, 79, 45, and 17% of vesicles, pustules, ulcers, and crusted lesions, respectively. Furthermore, we demonstrated, by fluctuation analysis of two encephalitis strains, that the ACVr variants were clonally distributed in the virus populations before exposure to ACV and did not result from rapid adaptation to ACV. Full text Full text is available as a scanned copy of the original print version. Virol. A three-dimensional reconstruction computed at 18-Å resolution from cryoelectron micrographs showed m100 procapsids to be structurally indistinguishable from procapsids assembled in vitro.

Zhu, S. Preliminary evidence using PAP staining indicates that the virus reaches the stroma at the same time as the epithelium, via the sensory nerves. Although no effective treatment exists for the eradication of genital herpes, several antiviral drugs are available to treat outbreak episodes and minimize disease symptoms. When both methods were used, HSV was identified in 97, 79, 45, and 17% of vesicles, pustules, ulcers, and crusted lesions, respectively. D. Challberg, E. In an effort to determine the requirement for oriL in latency, ts+7 was compared with wild-type virus for its ability to establish, maintain, and be reactivated from latent infection in a murine eye model.
isolation of a lacZ insertion mutant and association of the UL5 gene product with other members of the helicase-primase complex.

Like all herpesviruses, HSV-1 consists of an icosahedral capsid surrounded by a membrane envelope with the double-stranded DNA (dsDNA) genome contained inside the capsid. R. Lehman, Proc. Thus, the development of a novel class of anti-HSV agents with a mechanism of action that targets a viral protein essential for replication is desirable. Acad. Sci. USA 86:2186-2189, 1989).

The other members of the complex are viral proteins encoded by genes UL8 and UL52. The events of capsid formation prior to DNA encapsidation have been examined in insect cell extracts containing capsid proteins and in a purified system in which capsids are formed from purified proteins (24–26, 42). An insertion mutant containing a mutation in the UL5 gene (hr99) was isolated by using the insertional mutagen ICP6::lacZ, in which the Escherichia coli lacZ gene is expressed under control of the viral ICP6 promoter. When grown on Vero cells, hr99 does not form plaques or synthesize viral DNA, although both defects are complemented efficiently on the L2-5 cells. A signed informed consent form was obtained from each patient before the initiation of any study-specific procedure. Furthermore, since no detectable UL5 protein is synthesized in hr99-infected cells, these cells provide a valuable control not only for the detection of the UL5 protein itself but also for the detection of protein-protein interactions with UL8 and UL52 by coimmunoprecipitation. In addition, the lacZ insertion in hr99 provides a convenient screening system for the introduction of site-specific mutations into the viral genome (L.

Zhu and S. K. Electron micrographs of cells infected with HSV-1 ts1201 showed that the capsids accumulating at the nonpermissive temperature (NPT) were spherical like procapsids and sensitive to disassembly when the cells were incubated at 0°C. Virol. 66:469-479, 1992). The number of plaques present in each well was determined by counting under a microscope.

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