scholarly journals RATIOS OF VACCINIA VIRUS PARTICLES TO VIRUS INFECTIOUS UNITS

1959 ◽  
Vol 110 (3) ◽  
pp. 461-480 ◽  
Author(s):  
John R. Overman ◽  
D. Gordon Sharp
Keyword(s):  
Vaccinia Virus ◽  
Chorioallantoic Membrane ◽  
Growth Curves ◽  
Infective Virus ◽  
Ratio Increase ◽  
Marked Rise ◽  
Rabbit Skin ◽  
Virus Particles ◽  
Tissue Extracts ◽  
Particle Counts

Total virus particle counts, infectivity titrations and the ratios between particles and infective units have been determined for vaccinia virus infected tissues. Growth curves of vaccinia in the chorioallantoic membrane are characterized by relatively low ratios from 1 to 4 days after inoculation and a marked rise in the ratio at more prolonged intervals. Ratio determinations of vaccinia virus passages in the egg, rabbit skin, and guinea pig skin have been made to study the phenomenon of adaptation in different hosts. The embryonated egg chorioallantoic membrane shows no variation in the ratio of particles to infectious units during passage and it is concluded that this host is completely susceptible to vaccinia. During adaptive passages on the skin of rabbits and guinea pigs relatively large amounts of non-infective virus appear as indicated by a rise in the particle-infectivity ratios. The extent of ratio increase appears related to the general resistance of the host to the virus. Finally, treatment of crude tissue extracts with sonic vibration is described as an aid in dispersing the virus particles for quantitative particle counts.

scholarly journals The growth in vitro of vaccinia virus in chick embryo chorio-allantoic memberane, minced embryo and cell suspensions

1957 ◽  
Vol 55 (3) ◽  
pp. 347-360 ◽  
Author(s):  
H. B. Maitland ◽  
D. I. Magrath
Keyword(s):  
Chick Embryo ◽  
Vaccinia Virus ◽  
Growth Curve ◽  
Chorioallantoic Membrane ◽  
Growth Cycle ◽  
Cell Suspensions ◽  
Rabbit Skin ◽  
Chick Chorioallantoic Membrane ◽  
Considerable Doubt

The growth curve of rabbit skin-adapted vaccinia virus in the chick chorioallantoic membrane incubated in Hanks' solution showed a drop in titre of virus for about 10 hr. followed by growth. At least 25% of virus, sometimes more, remained infective. A similar fall in titre was observed in heated membranes in which the virus did not grow and this occurred also when membranes, either normal or heated, were infected and disintegrated before incubation.The growth curve of virus in minced chick-embryo was similar to that in chorioallantoic membrane.Virus in cell suspensions prepared from chick embryo and incubated in a nutrient medium showed only a small loss of infectivity before growth in some experiments and rarely dropped below 65–70 % of the original titre in others.These results throw considerable doubt on the view that loss of infectivity preceding growth of vaccinia virus should be interpreted as an essential part of a growth cycle.

scholarly journals ON THE ROLE OF RIBONUCLEIC ACID IN ANIMAL VIRUS SYNTHESIS

1960 ◽  
Vol 111 (3) ◽  
pp. 351-368 ◽  
Author(s):  
Igor Tamm ◽  
Rostom Bablanian
Keyword(s):  
Herpes Simplex ◽  
Vaccinia Virus ◽  
Ribonucleic Acid ◽  
Chorioallantoic Membrane ◽  
Host Cells ◽  
Active Inhibitor ◽  
Herpes Simplex Viruses ◽  
Virus Particles ◽  
Highly Active

Ribonuclease is a highly active inhibitor of vaccinia virus multiplication in vitro in the chorioallantoic membrane removed from embryonated chicken eggs. It is also a highly active inhibitor of pock formation by vaccinia and herpes simplex viruses on the chorioallantoic membrane in vivo. Marked inhibitory effects were obtained with 12.5 µg. of RNase. However, complete inhibition was not obtained with several hundred micrograms of the enzyme. RNase caused no inactivation of the infectivity of vaccinia virus particles but it had a marked inhibitory effect on multiplication of this virus when administered many hours after infection of host cells had occurred. RNase also failed to inactivate the infectivity of herpes simplex virus particles. The results obtained indicate that ribonucleic acid is necessary for the multiplication of two DNA-containing viruses; i.e., vaccinia and herpes simplex.

scholarly journals Human vaccinia-like virus outbreaks in São Paulo and Goiás States, Brazil: virus detection, isolation and identification

2004 ◽  
Vol 46 (6) ◽  
pp. 315-322 ◽  
Author(s):  
Teresa Keico Nagasse-Sugahara ◽  
Jonas José Kisielius ◽  
Marli Ueda-Ito ◽  
Suely Pires Curti ◽  
Cristina Adelaide Figueiredo ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Chorioallantoic Membrane ◽  
Virus Detection ◽  
Epidemiological Data ◽  
Sao Paulo ◽  
Cowpox Virus ◽  
São Paulo ◽  
Isolation And Identification ◽  
Virus Particles ◽  
The Face

Since October 2001, the Adolfo Lutz Institute has been receiving vesicular fluids and scab specimens of patients from Paraíba Valley region in the São Paulo and Minas Gerais States and from São Patricio Valley, in the Goiás State. Epidemiological data suggested that the outbreaks were caused by Cowpox virus or Vaccinia virus. Most of the patients are dairy milkers that had vesiculo-pustular lesions on the hands, arms, forearms, and some of them, on the face. Virus particles with orthopoxvirus morphology were detected by direct electron microscopy (DEM) in samples of 49 (66.21%) patients of a total of 74 analyzed. Viruses were isolated in Vero cell culture and on chorioallantoic membrane (CAM) of embryonated chicken eggs. Among 21 samples submitted to PCR using primers for hemagglutinin (HA) gene, 19 were positive. Restriction digestion with TaqI resulted in four characteristic Vaccinia virus fragments. HA nucleotide sequences showed 99.9% similarity with Cantagalo virus, described as a strain of Vaccinia virus. The only difference observed was the substitution of one nucleotide in the position 616 leading to change in one amino acid of the protein in the position 206. The phylogenetic analysis showed that the isolates clustered together with Cantagalo virus, other Vaccinia strains and Rabbitpox virus.

scholarly journals The vaccinia virus F12L protein is associated with intracellular enveloped virus particles and is required for their egress to the cell surface

2002 ◽  
Vol 83 (1) ◽  
pp. 195-207 ◽  
Author(s):  
Henriette van Eijl ◽  
Michael Hollinshead ◽  
Gaener Rodger ◽  
Wei-Hong Zhang ◽  
Geoffrey L. Smith
Keyword(s):  
Cell Surface ◽  
Vaccinia Virus ◽  
Virus Particles ◽  
Enveloped Virus ◽  
Virus Morphogenesis ◽  
C Terminus ◽  
Specific Proteins ◽  
Infected Cells ◽  
Mature Virus ◽  
Confocal And Electron Microscopy

The vaccinia virus (VV) F12L gene encodes a 65 kDa protein that is expressed late during infection and is important for plaque formation, EEV production and virulence. Here we have used a recombinant virus (vF12LHA) in which the F12L protein is tagged at the C terminus with an epitope recognized by a monoclonal antibody to determine the location of F12L in infected cells and whether it associates with virions. Using confocal and electron microscopy we show that the F12L protein is located on intracellular enveloped virus (IEV) particles, but is absent from immature virions (IV), intracellular mature virus (IMV) and cell-associated enveloped virus (CEV). In addition, F12L shows co-localization with endosomal compartments and microtubules. F12L did not co-localize with virions attached to actin tails, providing further evidence that actin tails are associated with CEV but not IEV particles. In vΔF12L-infected cells, virus morphogenesis was arrested after the formation of IEV particles, so that the movement of these virions to the cell surface was inhibited and CEV particles were not found. Previously, virus mutants lacking IEV- or EEV-specific proteins were either unable to make IEV particles (vΔF13L and vΔB5R), or were unable to form actin tails after formation of CEV particles (vΔA36R, vΔA33R, vΔA34R). The F12L deletion mutant therefore defines a new stage in the morphogenic pathway and the F12L protein is implicated as necessary for microtubule-mediated egress of IEV particles to the cell surface.

Multiplication of vaccinia virus in the chorioallantoic membrane in vitro

Virology ◽  
1957 ◽  
Vol 3 (1) ◽  
pp. 173-184 ◽  
Author(s):  
John R. Overman ◽  
Igor Tamm
Keyword(s):  
Vaccinia Virus ◽  
Chorioallantoic Membrane

scholarly journals Vaccinia Virus G1 Protein, a Predicted Metalloprotease, Is Essential for Morphogenesis of Infectious Virions but Not for Cleavage of Major Core Proteins

2004 ◽  
Vol 78 (13) ◽  
pp. 6855-6863 ◽  
Author(s):  
Camilo Ansarah-Sobrinho ◽  
Bernard Moss
Keyword(s):  
Vaccinia Virus ◽  
Virus Replication ◽  
Protein A ◽  
Proteolytic Processing ◽  
Wild Type Virus ◽  
Sequence Information ◽  
Wild Type ◽  
Active Site Residues ◽  
Virus Particles ◽  
Core Proteins

ABSTRACT Genes encoding orthologs of the vaccinia virus G1 protein are present in all poxviruses for which sequence information is available, yet neither the role of the protein nor its requirement for virus replication is known. G1 was predicted to be involved in the cleavage of core proteins, based on a transfection study and the presence of an HXXEH motif found in a subset of metallopeptidases. In the present study, we engineered a recombinant vaccinia virus containing a single copy of the G1L gene with a C-terminal epitope tag that is stringently regulated by the Escherichia coli lac repressor. In the absence of inducer, expression of G1 was repressed and virus replication was inhibited. Rescue of infectious virus was achieved by expression of wild-type G1 in trans, but not when the putative protease active site residues histidine-41, glutamate-44, or histidine-45 were mutated. Nevertheless, the synthesis and proteolytic processing of major core and membrane proteins appeared unaffected under nonpermissive conditions, distinguishing the phenotype of the G1L mutant from one in which the gene encoding the I7 protease was repressed. Noninfectious virus particles, assembled in the absence of inducer, did not attain the oval shape or characteristic core structure of mature virions. The polypeptide composition of these particles, however, closely resembled that of wild-type virus. Full-length and shorter forms of the G1 protein were found in the core fraction of virus particles assembled in the presence of inducer, suggesting that G1 is processed by self-cleavage or by another protease.

MORPHOLOGICAL DEVELOPMENT OF CHIKUNGUNYA VIRUS

1966 ◽  
Vol 12 (5) ◽  
pp. 895-900 ◽  
Author(s):  
Marybelle M. T. Chain ◽  
Frances W. Doane ◽  
D. M. McLean
Keyword(s):  
Chikungunya Virus ◽  
Phosphotungstic Acid ◽  
Morphological Development ◽  
Infective Virus ◽  
Chick Embryo Fibroblast ◽  
Virus Particles ◽  
Outer Membranes ◽  
Extracellular Virus ◽  
Dense Cores ◽  
Infected Cells

Chikungunya virus was first detected by electron microscopy of primary chick embryo fibroblast cultures 5 hours after inoculation. Two types of particles were observed in the cytoplasm of infected cells, but not in uninoculated cells. The smaller, presumably precursor particles, measured 260 to 280 Å. The larger particles contained dense cores 260 to 280 Å in diameter and outer shells 500 to 560 Å in diameter. In cell lysates stained negatively with phosphotungstic acid 12 and 24 hours after inoculation, virus particles surrounded by fine outer membranes showed diameters of 540 to 580 Å. Infective virus was first detected 5 hours after inoculation and maximum yields of cell-associated and extracellular virus were attained at 8 to 10 hours.

Completely Dispersed Suspensions of Vaccinia Virus Particles.

1964 ◽  
Vol 117 (1) ◽  
pp. 101-105 ◽  
Author(s):  
G. J. Galasso ◽  
D. G. Sharp
Keyword(s):  
Vaccinia Virus ◽  
Virus Particles
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