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DNA from Cotia virus, an unclassified poxvirus, was mapped by overlapping fragment analysis using the restriction endonucleases HindIII, Pst I, BamHI, XhoI, SalI, and SmaI. The linear genome was 177 kbp in length and possessed inverted terminal repeats and cross-links. A Cotia virus thymidine kinase (TK) gene was detected and mapped to about 74 kbp from the left end of the genome using degenerate...
The complete nucleotide sequences of five double-stranded RNA genome segments of the Diadromus pulchellus reovirus (DpRV) have been determined. They consist of 985, 1240, 1318, 1652, and 4230 bp. Each segment contains at least one putative open reading frame encoding 33-, 40-, 45-, 49-, and 148-kDa proteins, respectively. The proteins have no significant similarities with sequences in data banks....
Core particles of bluetongue virus serotype 1 (South Africa) have been crystallized. The crystals, which grow up to 0.8 mm in diameter, belong to a primitive orthorhombic space group and have point group symmetry 222. The unit cell dimensions are 754 x 796 x 823 Å 3 and the crystallographic asymmetric unit contains one-half of a core particle. The best crystals diffract strongly to 4.8 Å Bragg...
The mode and tempo of arbovirus evolution and dispersal can help to explain the dynamics of pandemics, viral outbreaks, and emerging viruses. By comparing nucleotide and deduced amino acid sequences of their envelope proteins, we describe the continuous distribution of the tick-borne encephalitis (TBE) complex viruses, the most important flaviviruses in Europe, across major geographical areas and...
Partial-length gene 4 cDNA encompassing the VP8* portion of VP4 from chicken rotavirus (RV) Ch-1, turkey RV Ty-1 and Ty-3, and pigeon RV PO-13 was sequenced and compared to RV 993/83 isolated from a calf with diarrhea. Ninety-six percent amino acid sequence identity was seen between VP8* from calf RV 993/83 and pigeon RV PO-13, while only 77 to 83% identity was seen in comparison with turkey and chicken...
Virus isolates resistant to 1-[(2-hydroxyethoxy) methyl]-6-(phenylthio)thymine (HEPT) and a highly potent HEPT derivative, [1-benzyloxymethyl-5-ethyl-6-(α-pyridylthio)uracil] (NSC 648400, E-BPTU), were selected in cell culture. Cross-resistance evaluation indicated that the two drug resistant virus isolates were phenotypically distinct from one another although each of the virus isolates was resistant...
To examine whether the mutation of protease in an HIV-1 resistant to a protease inhibitor affects the virus phenotype in vitro, the infectivity of the protease inhibitor-escape-virus was compared to that of the parent virus, In different T-cell lines, the infectivity of the escape virus was impaired by 10-fold compared to the parent virus. MT-4 cell killing by the escape virus, measured using the...
A chimeric gene encoding a fusion protein consisting of the DNA-binding domain of the immediate-early (IE) protein of pseudorabies virus (PRV) and a tail-truncated Vmw65 of herpes simplex virus 1, lacking the transcription activation domain, was constructed The chimeric gene product inhibited transcription from the PRV IE promoter in a transient expression assay. A HeLa cell line stably transformed...
Influenza virus hemagglutinin (HA) subtype H 7 expressed from a baculovirus vector in insect cells requires cysteine residues for palmitoylation. Mutant HA devoid of fatty acids shows hemagglutinating and hemolytic activities almost identical to those of the acylated wild-type HA (wt). Using a membrane mixing assay (R 18 ), neither the kinetics nor the pH dependence of fusion...
Cytotoxic T lymphocytes (CTL) play a pivotal role in preventing persistent viral infections and aborting acute infections. H-2D b -restricted CTL optimally recognize a specific peptide of 9 to 11 amino acids (aa) derived from a viral protein and held in place (restricted) by a MHC class I glycoprotein on the surfaces of infected cells. Only three peptide sequences with the appropriate D ...
The vaccinia virus/bacteriophage T7 hybrid transient expression system employs a recombinant vaccinia virus that encodes the T7 RNA polymerase gene, a plasmid vector with a gene of interest regulated by a T7 promoter, and any cell line suitable for infection and transfection. Although high expression in a majority of cells is achieved. the severe cytopathic effects of vaccinia virus and the safety...
Defective interfering (DI) influenza virus A/WSN (H1N1) grown in embryonated eggs protected adult mice from a lethal respiratory infection with A/WSN virus. Eighteen bands of putative DI RNA, ranging in size from about 230 to 1020 nt, were identified in this preparation by reverse transcription and polymerase chain reaction using a segment-specific 3' primer and a segment-universal 5' primer. Every...
The relative prevalence of Sendai virus-specific CD4 + T helper precursors (Thp) has been assessed by limiting dilution analysis (LDA). Within 10 days after intranasal inoculation of C57BL/6 mice, Thp prevalence increased from <1 IN 10 5 to approximately 1 in 200 in CD4 + T-cell-enriched spleen and mediastinal lymph node populations. These frequencies remained elevated...
Central nervous system (CNS) disorders are frequent in HIV-1-infected individuals, particularly in newborns and children, and are accompanied by histological alterations resulting in neuronal loss. Although several tumor-derived neuroectodermal cell lines can be infected by HIV-1, it has been reported that primary neural cells cannot be infected after they differentiate. However, pediatric AIDS is...
An element(s) within a 386-nucleotide segment (TCV-386 RNA) of the turnip crinkle virus (TCV) genome was previously implicated in virus assembly nucleation. To localize the proposed high-affinity binding determinants, we analyzed the ability of the coat protein to bind the full-length and truncated derivatives of the TCV-386 RNA using gel retardation and nitrocellulose filter retention assays. Quantitation...
The tobacco etch virus 27-kDa nuclear inclusion a (NIa) proteinase was expressed in Escherichia coli as a recombinant fusion protein containing a seven histidine tag at the amino-terminus. Catalytically active and inactive (by virtue of a single amino acid change) forms of the proteinase were purified to homogeneity in a two-column chromatographic procedure The active form of the proteinase was slowly...
A quantitative ribonuclease protection assay (RPA) was developed in order to rapidly and accurately measure the levels and timing of latency-associated transcript (LAT) expression in ganglia latently infected with wild-type and mutant herpes simplex virus (HSV). Use of this assay in parallel with measurement of viral titers in murine trigeminal ganglia demonstrated that the peak of viral replication...
Passage of cucumber necrosis virus (CNV) containing defective interfering (DI) RNAs through cucumber plants decreased the accumulation of DI RNAs to undetectable levels. Subsequent passages in two Nicotiana species (Nicotiana benthamiana or N. clevelandii) resulted in the appearance of DI RNA species that were larger than the DI RNAs observed during exclusive serial passages of CNV through the Nicotiana...
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