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High concentrations of DIP lead to a state of persistent infection in which infected cells become lysis resistant but release both standard virus and DIP (Henry et al., 1979, 1980; Robinson et al., 1980; Dauenhauer et al., 1982). Taken together, data from this study clearly demonstrates that sequential cell culture passage of the neuropathogenic T953 strain of EHV-1 results in attenuation for young adult BALB/C mice. In transient transfection assays, the IR2P down-regulated luciferase reporter activity that was under the control of the sole IE promoter or early viral promoters such as EICP0, TK, IR4, and UL5 in a dose-dependent manner (Kim et al., 2006). However, while HSV-1 is highly prevalent in the human population, in the vast majority of cases giving no obvious signs of disease, for use as a vector, the virus must be disabled for safety and to minimize toxicity to target cells. High concentrations of DIP lead to a state of persistent infection in which infected cells become lysis resistant but release both standard virus and DIP (Henry et al., 1979, 1980; Robinson et al., 1980; Dauenhauer et al., 1982). This requirement does not appear to stem from a need for local initiation of DNA synthesis, since deletion of the two origins that lie within HSV-1 inverted repeats does not alter their inversion efficiency (54). Introduction Bluetongue virus (BTV), the prototype of the genus Orbivirus within the family Reoviridae, is the causative agent of bluetongue disease in many species of domestic ruminants, especially sheep.

Infectious laryngotracheitis is a contagious respiratory disease of chickens which causes severe losses in the poultry industry (1). In addition, areas of heterogeneity were detected at the L terminus, within the S segment, and at a split variable locus in the L region. J. Strain Wh shows much higher identity to EHV-8 than to other alphaherpesviruses based on the amino acid sequence deduced from the glycoprotein G gene. The transposon library is a resource for comprehensive functional analysis of the HVS25A genome, with multiple mutants available in any of the predicted genes of EHV-1. These references are in PubMed. In contrast, a 1.7-kb mRNA appears at later times postinfection and is designated as a gamma-1 transcript, since its synthesis is significantly reduced by phosphonoacetic acid.

This may not be the complete list of references from this article. RNA mapping data revealed that IR4 has two promoters that are regulated differentially during a lytic infection. To identify the IR6 protein and ascertain its properties, we generated an IR6-specific polyclonal antiserum to a TrpE/IR6 fusion protein containing 129 amino acids (residues 134 to 262) of the IR6 protein. A site was used in gel shift analysis to show that the EHV-1 origin-binding protein bound to the same consensus site as the HSV-1 origin-binding protein, 5′-CGTTCGCACTT-3′. In addition, infection of RK13 cells with ETIF-deleted or parental virus at an MOI of 1 yielded comparable amounts of Gag and GFP, indicating that neither gene delivery nor transgene expression is affected by the deletion of the ETIF gene (Fig. The miniF plasmid pHA2 (hatched area) (1) was inserted (46) into the gene locus of gene 71. Furthermore, comparison of known origin sequences demonstrated that a 9-bp sequence, CGTTCGCAC, which is conserved among all origins of replication of human lytic herpesviruses and which is contained within the 18-bp region in herpes simplex virus type 1 origins shown by others to be protected by an origin-binding protein (P.

Robinson, and D. These findings may contribute to defining the role of the UL31P single-stranded DNA-binding protein in EHV-1 DNA replication. Gel shift assays revealed that the glutathione S-transferase (GST)-IEQ495E(407-615) and GST-IEK498E(407-615) proteins failed to bind to the IE promoter, indicating that the Gln and Lys residues are important for the DNA binding activity. In addition, areas of heterogeneity were detected at the L terminus, within the S segment, and at a split variable locus in the L region. After in vitro transcription and translation, both constructs yielded a comigrating 23-kDa protein, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Two strains could be differentiated using BamHI restriction and were assigned to the EHV-1 1B prototype group. Serial, high multiplicity passage of equine herpesvirus 1 (EHV-1) leads to the generation of defective interfering particles (DIP).

The 151,601-nt genome encodes 76 distinct genes like other equine alphaherpesviruses, but genetically, EHV-3 is significantly more divergent. Specifically, STD EHV-1 DNA fragments encompassing the genomic regions present in DI particle DNA were inserted into the vector pAT153, and individual clones were tested by transfection assays for the ability to support the amplification and replication of plasmid DNA in EHV-1-infected cells. Serial, high multiplicity passage of equine herpesvirus 1 (EHV-1) leads to the generation of defective interfering particles (DIP). The IR2P was first detected in the nucleus at 5 h postinfection in equine herpesvirus 1 (EHV-1)-infected HeLa and equine NBL6 cells.