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Isolation and Characterization of a Herpes Simplex Virus Type 1 Mutant Containing a Deletion within the Gene Encoding the Immediate Early Polypeptide Vmw110

作者:N. D. Stow, E. C. Stow · 发表于:Journal of General Virology · 年份:1986 · DOI:10.1099/0022-1317-67-12-2571 · 被引用次数:424 · 研究领域:Herpesvirus Infections and Treatments、Virus-based gene therapy research、T-cell and Retrovirus Studies

Transfection experiments with plasmids containing immediate early (IE) genes of herpes simplex virus type 1 (HSV-1) have previously demonstrated a role for the IE polypeptide Vmw110 (ICP0) in stimulating expression from plasmid-encoded early gene promoters. To gain further insights into the function of Vmw110 we isolated a deletion mutant specifying a truncated form of the polypeptide which had been shown to be inactive in transfection assays. This mutant, dl1403, contained a 2 kb deletion within both the TRL and IRL copies of the Vmw110 gene, and encoded a polypeptide consisting of the original N-terminal 105 amino acids followed by 56 amino acids specified by a reading frame not used by Vmw110. dl1403 was able to replicate and produce plaques on baby hamster kidney (BHK) cells but the yield of infectious virus was 20- to 100-fold lower than obtained with wild-type HSV-1. Surprisingly, comparison of polypeptide synthesis, DNA replication and DNA encapsidation in cells infected with 5 p.f.u./cell dl1403 or wild-type HSV-1 revealed no significant differences. In addition similar numbers of particles were produced in cells infected with the two viruses, resulting in stocks of dl1403 exhibiting significantly higher particle/p.f.u. ratios. The efficiency of plaquing of dl1403 was greatly reduced in Vero and human foetal lung cells compared with BHK cells, but following infection with 5 p.f.u./cell similar yields of infectious virus were obtained from all three cell lines. Marker ...