- Volume 66, Issue 3, 1985
Volume 66, Issue 3, 1985
- Animal
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Coding Sequence of Coronavirus MHV-JHM mRNA 4
More LessSUMMARYA coding sequence at the 5′ end of mRNA 4 of the coronavirus MHV-JHM was determined by M13/chain-terminator sequencing of cloned cDNA. An open reading frame of 417 bases with the potential to encode a polypeptide of mol. wt. 15200 (139 residues) was identified. The 3′ end of the open reading frame overlapped by 16 bases the start of an open reading frame found in mRNA 5. The translation product of mRNA 4 was predicted to be a basic polypeptide rich in threonine. It had a large hydrophobic region near the amino terminus and a basic carboxy terminus. An intracellular, virus-specific polypeptide, which has been previously described as having a mol. wt. of 14000 to 14500 has the size and charge characteristics of such a translation product.
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Peptide Mapping of Envelope-related Glycoproteins Specified by the Flaviviruses Kunjin and West Nile
More LessSUMMARYGlycoproteins detected in Vero cells infected by the flaviviruses West Nile and Kunjin were examined by gel electrophoresis and peptide mapping. Two major glycoproteins, gp66 and gp54, were observed in West Nile virus-infected cells labelled for short time periods with [3H]mannose. A third glycoprotein, gp58, was present in smaller amounts. Pulse-labelling experiments suggested that gp66 was a precursor of gp54. Peptide mapping of [3H]leucine-labelled gp66, gp54 and the envelope glycoprotein E of West Nile virus demonstrated that gp66 and gp54 were related to E, and that the peptides of gp54 were a subset of those of gp66. Peptide mapping of the corresponding Kunjin virus-specified glycoproteins (gp66, gp59 and gp53) showed that the [3H]leucine-labelled peptides of gp53 and gp59 were similar and were contained within gp66. Since we have shown previously that gp59 and gp53 are related to E of Kunjin virus, we conclude that cells infected by West Nile or Kunjin viruses contain a similar set of E-related glycoproteins.
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Lack of Quantitative Correlation between the Neutralization of Poliovirus and the Antibody-mediated pI Shift of the Virions
More LessSUMMARYThe effect of mono- and polyclonal antibodies on the infectivity and pI (isoelectric pH) of type 1 poliovirus was studied. According to Mandel’s hypothesis, the isoelectric pH of poliovirus should change to about pI 4 upon neutralization. However, several antibodies did not follow this rule. Moreover, when antibodies did shift the pI, no quantitative correlation existed between the extent of neutralization and the amount of poliovirus shifted to low pI.
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Human Papillomavirus Type 16 Recombinant DNA Is Maintained as an Autonomously Replicating Episome in Monkey Kidney Cells
More LessSUMMARYHuman papillomavirus type 16 (HPV16) DNA cloned in the expression vector pSV2-neo has been shown to be maintained in transfected monkey kidney cells as an autonomously replicating episome at a level of 2 to 10 copies per cell. Integration of pSV2-neo/HPV16 DNA with the host genome occurred in transfected mouse fibroblasts. Neither type of cell appeared to be phenotypically transformed.
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Recombinants between Vaccinia and Ectromelia Viruses Bearing the Specific Pathogenicity Markers of Both Parents
More LessSUMMARYNineteen recombinants between vaccinia virus (VV) DNA− temperature-sensitive mutants and ectromelia virus (EMV) were characterized with respect to their biological properties and genome structure. Four of these recombinants acquired the pathogenicity for mice characteristic of EMV, while the pathogenicity for rabbits characteristic of VV was not only preserved, but even enhanced. Most unexpectedly, these recombinants with ‘double pathogenicity’, as well as four other recombinants, acquired a stable genetic trait which was not typical of the parental viruses, i.e. the ability to form haemorrhagic lesions on the chorioallantoic membrane of chick embryos. Approximate mapping of the genomes of these recombinants with restriction endonucleases showed that their DNA contained mostly VV sequences with a single detected insert of EMV DNA. In the ‘double pathogenicity’ recombinants, this insert was located in the central part of the genome, and its minimal size was about 16 Mdal.
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Small RNA Viruses Co-infecting the Pine Emperor Moth (Nudaurelia cytherea capensis)
More LessSUMMARYLarvae of the pine emperor moth, Nudaurelia cytherea capensis, naturally infected with Nudaurelia β virus (NβV), contained a second serologically unrelated virus which we have called Nudaurelia ω virus (NωV). NωV had a buoyant density of 1.285 g/ml in CsCl and yielded a single major polypeptide of 65000 daltons on gel electrophoresis. The particles of NωV were morphologically distinguishable from those of Nudaurelia viruses described by others, and tryptic peptide analyses indicated that NωV protein was distinct from that of NβV and NɛV. Both NωV and NβV contained a small stable fraction of particles with buoyant densities of about 1.33 g/ml.
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Effect of Heat Shock on Epstein–Barr Virus and Cytomegalovirus Expression
More LessSUMMARYThe effect of heat shock was investigated on lymphoblastoid cell lines Raji and P3HR1 harbouring Epstein-Barr virus (EBV) genomes, and on Vero cells abortively infected with human cytomegalovirus (HCMV). A heat shock at 44 °C for 10 min induced the appearance of EBV early antigens in Raji cells and increased the percentage of cells expressing EBV viral capsid antigens in P3HR1 cells. Heat shock performed on Vero cells just before HCMV infection resulted in an approximately fourfold increase in the number of cells exhibiting early nuclear antigens, and in the appearance of HCMV-induced Fc receptors.
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Comparative Electrophoretic Study of Polypeptides of Influenza A/H3N2 Viruses Isolated in Circumscribed Geographical Areas
More LessSUMMARYTwo distinct groups of influenza A/H3N2 viruses, closely related to A/Bangkok/1/79 and to A/Belgium/2/81, have been chosen from viruses isolated in Italy during 1981 to 1983 with the aim of analysing the biochemical composition of their polypeptides. The strains of each group have shown differences in electrophoretic migration rates in one or more proteins in comparison to the prototype viruses. Polypeptide mobility variations among isolates from circumscribed geographical areas and from single outbreaks have also been observed. In particular, there was a high degree of variability in the NS1 protein. The detection of biochemical differences among identical antigenic variants, probably the result of point mutations in polypeptide sequences or of genetic reassortment among different co-circulating human viruses, is a further expression of the peculiar ability of the influenza A virus to exhibit variation in internal proteins during its circulation.
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Extracellular Release of Enveloped Vaccinia Virus from Mouse Nasal Epithelial Cells in vivo
More LessSUMMARYThe release of vaccinia virus from mouse nasal epithelial cells infected in vivo was studied by electron microscopy. Intracellular naked vaccinia virus was enwrapped by Golgi membranes to form a double membrane intermediate. The outer membrane of the intermediate presumably fused with the plasma membrane, releasing extracellular enveloped virus. No signs of simple naked virus budding at the plasma membrane were observed. The majority of extracellular virus was enveloped and not naked.
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Polyethylene Glycol-mediated Infection of Cowpea Protoplasts with Sonchus Yellow Net Virus
More LessSUMMARYThe conditions favouring the infection of cowpea mesophyll protoplasts by Sonchus yellow net virus (SYNV) were determined. When 3 × 106 protoplasts were inoculated with 60 µg SYNV in 40% polyethylene glycol, 3 mm-CaCl2 at room temperature, over 90% of the surviving protoplasts became infected. Infectivity tests showed that the virus could be detected 12 h after inoculation.
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Cauliflower Mosaic Virus DNA Persists as Supercoiled Forms in Cultured Turnip Cells
More LessSUMMARYExplants taken from turnip leaves infected systemically with cauliflower mosaic virus (CaMV) proliferate callus tissue when cultured on an appropriate medium. CaMV DNA, detected by spot hybridization, was found to persist in these cultured cells for at least 45 days. Analysis of unencapsidated virus DNA by blot hybridization after agarose gel electrophoresis showed that the truncated DNA forms characteristic of infected leaf tissue were virtually absent from callus cells. These cells contained predominantly genome-length supercoiled CaMV DNA together with discrete subgenomic supercoiled forms.
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