scholarly journals ONE-STEP GROWTH CURVE OF WESTERN EQUINE ENCEPHALOMYELITIS VIRUS ON CHICKEN EMBRYO CELLS GROWN IN VITRO AND ANALYSIS OF VIRUS YIELDS FROM SINGLE CELLS

1954 ◽  
Vol 99 (2) ◽  
pp. 183-199 ◽  
Author(s):  
R. Dulbecco ◽  
Marguerite Vogt
Keyword(s):  
Chicken Embryo ◽  
Cell Layer ◽  
Growth Curves ◽  
Virus Growth ◽  
Embryo Cells ◽  
Cell Layers ◽  
A Cell ◽  
One Step ◽  
Step Growth

The rate of adsorption of WEE virus onto chicken embryo cells in vitro was determined both on a cell layer and on a cell suspension. One-step growth curves were determined in cell suspensions and on cell layers. The latent period varied between 2 and 3½ hours; it was shorter on cell layers and decreased with higher multiplicity of infection. The shortest period is probably the real latent period. The growth curves of the virus showed an initial exponential rise and reached a maximal constant value after 6 to 8 hours. The maximum virus yield per cell varied between 200 and 1000 on the cell layer, and between 100 and 200 in suspended cells. The yield of single infected cells was determined. An analysis of the distributions of the individual yields obtained after various periods of virus growth led to two main conclusions: (1) that virus is released from the same cell over a long period of time; (2) that one phase of the intracellular virus growth is exponential.

scholarly journals ONE-STEP GROWTH CURVES OF VARIOUS STRAINS OF INFLUENZA A AND B VIRUSES AND THEIR INHIBITION BY INACTIVATED VIRUS OF THE HOMOLOGOUS TYPE

1949 ◽  
Vol 89 (3) ◽  
pp. 279-285 ◽  
Author(s):  
Werner Henle ◽  
Evelyn B. Rosenberg
Keyword(s):  
Influenza A ◽  
Swine Influenza Virus ◽  
Swine Influenza ◽  
Growth Curves ◽  
Influenza B Virus ◽  
Influenza B ◽  
Virus Growth ◽  
One Step ◽  
Infected Cells ◽  
Step Growth

One-step growth curves of five strains of influenza A, one strain of swine influenza, and three strains of influenza B virus have been analyzed. The influenza A and swine influenza strains showed constant periods of 5 to 6 hours before newly formed virus was liberated from the infected cells, whereas 8 to 10 hours elapsed in the case of the influenza B strains. The yield of virus in the allantoic fluids, i.e. the number of ID50 released for every ID50 of seed virus adsorbed, was consistently higher in the case of the influenza A and swine influenza strains than in that of the influenza B viruses. Interruption of the cycle by injection of inactivated virus subsequent to infection can be achieved by any of the strains of the homologous type. However, cross-tests between influenza A and swine influenza virus led only to partial inhibition of virus growth.

Fiber1, but not fiber2, is the essential fiber gene for fowl adenovirus 4 (FAdV-4)

2021 ◽  
Author(s):  
Xiaohui Zou ◽  
Yejing Rong ◽  
Xiaojuan Guo ◽  
Wenzhe Hou ◽  
Bingyu Yan ◽  
...  
Keyword(s):  
Reverse Genetics ◽  
Growth Curves ◽  
Dependent Manner ◽  
Helper Plasmid ◽  
Fowl Adenovirus ◽  
Virus Adsorption ◽  
One Step ◽  
Step Growth ◽  
Lmh Cells

Fibre is the viral protein that mediates the attachment and infection of adenovirus to the host cell. Fowl adenovirus 4 (FAdV-4) possesses two different fibre trimers on each penton capsomere, and roles of the separate fibres remain elusive. Here, we attempted to investigate the function of FAdV-4 fibres by using reverse genetics approaches. Adenoviral plasmids carrying fiber1 or fiber2 mutant genes were constructed and used to transfect chicken LMH cells. Fiber1-mutated recombinant virus could not be rescued. Such defective phenotype was complemented when a fiber1-bearing helper plasmid was included for co-transfection. The infection of fiber-intact FAdV-4 (FAdV4-GFP) to LMH cells could be blocked with purified fiber1 knob protein in a dose-dependent manner, while purifed fiber2 knob had no such function. On the contrary, fiber2-mutated FAdV-4, FAdV4XF2-GFP, was successfully rescued. The results of one-step growth curves showed that proliferative capacity of FAdV4XF2-GFP was 10 times lower than that of the control FAdV4-GFP. FAdV4XF2-GFP also caused fewer deaths of infected chicken embryos than FAdV4-GFP did, which resulted from poorer virus replication in vivo. These data illustrated that fiber1 mediated virus adsorption and was essential for FAdV-4, while fiber2 was dispensable although it significantly contributed to the virulence.

Effectiveness of exogenous DNA transfer to chicken embryo cells in vitro and in vivo using retroviral vectors

2007 ◽  
Vol 33 (3) ◽  
pp. 180-182 ◽  
Author(s):  
N. A. Volkova ◽  
A. O. Tulyakova ◽  
L. A. Volkova ◽  
N. A. Zinov’eva ◽  
L. K. Ernst ◽  
...  
Keyword(s):  
Chicken Embryo ◽  
Retroviral Vectors ◽  
Dna Transfer ◽  
Exogenous Dna ◽  
Embryo Cells

scholarly journals Random Transposon-Mediated Mutagenesis of the Essential Large Tegument Protein pUL36 of Pseudorabies Virus

2010 ◽  
Vol 84 (16) ◽  
pp. 8153-8162 ◽  
Author(s):  
Britta S. Möhl ◽  
Sindy Böttcher ◽  
Harald Granzow ◽  
Walter Fuchs ◽  
Barbara G. Klupp ◽  
...  
Keyword(s):  
Amino Acids ◽  
Pseudorabies Virus ◽  
Plaque Assay ◽  
Tegument Protein ◽  
Nuclear Pore ◽  
Viral Spread ◽  
One Step ◽  
Step Growth ◽  
Insertion Mutants

ABSTRACT Homologs of the pseudorabies virus (PrV) essential large tegument protein pUL36 are conserved throughout the Herpesviridae. pUL36 functions during transport of the nucleocapsid to and docking at the nuclear pore as well as during virion formation after nuclear egress in the cytoplasm. Deletion analyses revealed several nonessential regions within the 3,084-amino-acid PrV pUL36 (S. Böttcher, B. G. Klupp, H. Granzow, W. Fuchs, K. Michael, and T. C. Mettenleiter, J. Virol. 80:9910-9915, 2006; S. Böttcher, H. Granzow, C. Maresch, B. Möhl, B. G. Klupp, and T. C. Mettenleiter, J. Virol. 81:13403-13411, 2007), while the C-terminal 62 amino acids are essential for virus replication (K. Coller, J. Lee, A. Ueda, and G. Smith, J. Virol. 81:11790-11797, 2007). To identify additional functional domains, we performed random mutagenesis of PrV pUL36 by transposon-mediated insertion of a 15-bp linker. By this approach, 26 pUL36 insertion mutants were selected and tested in transient transfection assays for their ability to complement one-step growth and/or viral spread of a PrV UL36 null mutant. Ten insertion mutants in the N-terminal half and 10 in the C terminus complemented both, whereas six insertion mutants clustering in the center of the protein did not complement in either assay. Interestingly, several insertions within conserved parts yielded positive complementation, including those located within the essential C-terminal 62 amino acids. For 15 mutants that mediated productive replication, stable virus recombinants were isolated and further characterized by plaque assay, in vitro growth analysis, and electron microscopy. Except for three mutant viruses, most insertion mutants replicated like wild-type PrV. Two insertion mutants, at amino acids (aa) 597 and 689, were impaired in one-step growth and viral spread and exhibited a defect in virion maturation in the cytoplasm. In contrast, one functional insertion (aa 1800) in a region which otherwise yielded only nonfunctional insertion mutants was impaired in viral spread but not in one-step growth without a distinctive ultrastructural phenotype. In summary, these studies extend and refine previous analyses of PrV pUL36 and demonstrate the different sensitivities of different regions of the protein to functional loss by insertion.

scholarly journals PHAGE FORMATION IN STAPHYLOCOCCUS MUSCAE CULTURES

1950 ◽  
Vol 34 (2) ◽  
pp. 231-250 ◽  
Author(s):  
Winston H. Price
Keyword(s):  
Aspartic Acid ◽  
Burst Size ◽  
Synthetic Medium ◽  
Growth Curves ◽  
Acid Synthesis ◽  
Virus Protein ◽  
Complete Medium ◽  
Multiplication Rate ◽  
One Step ◽  
Step Growth

1. Four strains of Staphylococcus muscae have been isolated which differ in their growth rates and phage syntheses in Fildes' synthetic medium. 2. Two of the strains when singly infected cannot release phage in Fildes' synthetic medium unless a substance present in certain acid-hydrolyzed proteins is added to the medium. One of these strains also requires other substance(s) present in acid-hydrolyzed proteins in order to grow in Fildes' medium. 3. The two strains which do not require the addition of the phage-stimulating factor have been found either to synthesize this substance, or one similar to it. One of these strains will not grow in Fildes' medium unless substance(s) present in acid-hydrolyzed proteins is added to the medium. 4. The purified acid-hydrolyzed protein factor necessary for virus liberation does not affect the multiplication rate of uninfected S. muscae cells in Fildes' synthetic medium. 5. The substance is not needed for the adsorption or the invasion of the host cell by the virus. In the absence of the factor, the virus is adsorbed to the cell and "kills" it. 6. An analysis carried out by means of the one-step growth curve technique has indicated that the substance is not concerned simply with the mechanism of virus release, but is necessary for some initial stage in virus synthesis. 7. With one bacterial strain not requiring the AHPF, aspartic acid had to be present at least during the minimum latent period for the cell to form virus. 8. In the absence of aspartic acid, the virus was adsorbed to the cell and killed it, but no virus was released from singly infected bacteria. 9. If the cells were grown in a medium containing aspartic acid and then resuspended in the medium minus aspartic acid, no virus was released, although such cells contained at least two times the amount of aspartic acid necessary for the burst size in the complete medium. 10. Aspartic acid, a constituent of the virus particle, appears from an analysis of one-step growth curves to take part in the initial phase of phage synthesis. 11. The effect of amino acids on virus formation is discussed in relation to the time sequence of virus protein and desoxyribonucleic acid synthesis.

STUDIES OF INFECTIOUS BOVINE RHINOTRACHEITIS VIRUS: 1. PLAQUE ASSAY AND SOME CHARACTERISTICS IN BOVINE KIDNEY CELLS

1963 ◽  
Vol 9 (4) ◽  
pp. 567-576 ◽  
Author(s):  
Leslie R. Sabina ◽  
Raymond C. Parker
Keyword(s):  
Infectious Virus ◽  
Plaque Assay ◽  
Growth Curves ◽  
Kidney Cells ◽  
Infectious Bovine Rhinotracheitis ◽  
High Input ◽  
Bovine Kidney ◽  
One Step ◽  
Step Growth ◽  
Infectious Bovine Rhinotracheitis Virus

A reproducible plaquing procedure for infectious bovine rhinotracheitis virus (IBRV) in an established bovine kidney cell line is reported. The validity of this system for quantitative analysis has been established by conventional methods.After infection at different multiplicities, one-step growth curves have shown that the eclipse period for IBRV lasts approximately 4 hours and that the infectious virus increases at a logarithmic rate for 12 to 14 hours. The virus yield with the low and high input is 30 PFU and 210 PFU per cell, respectively. Only 1 to 9% of the total virus is released at 24 hours postinfection. The data presented indicate the half-life of IBRV at 37 °C and 42 °C to be 16 and 3.5 hours, respectively. A comparison of hyperimmune bovine and rabbit sera has shown that 92% of the infective particles are neutralized within 30 minutes.

scholarly journals Rethinking the Evolution of Single-Stranded RNA (ssRNA) Bacteriophages Based on Genomic Sequences and Characterizations of Two R-Plasmid-Dependent ssRNA Phages, C-1 and Hgal1

2012 ◽  
Vol 194 (18) ◽  
pp. 5073-5079 ◽  
Author(s):  
Sherin Kannoly ◽  
Yongping Shao ◽  
Ing-Nang Wang
Keyword(s):  
Genome Organization ◽  
Cell Attachment ◽  
Phylogenetic Analyses ◽  
Bacterial Species ◽  
Growth Curves ◽  
Conjugative Plasmid ◽  
Content Type ◽  
Conjugative Plasmids ◽  
One Step ◽  
Step Growth

ABSTRACTWe have sequenced and characterized two R-plasmid-dependent single-stranded RNA bacteriophages (RPD ssRNA phages), C-1 and Hagl1. Phage C-1 requires a conjugative plasmid of the IncC group, while Hgal1 requires the IncH group. Both the adsorption rate constants and one-step growth curves are determined for both phages. We also empirically confirmed the lysis function of the predicted lysis genes. Genomic sequencing and phylogenetic analyses showed that both phages belong to theLevivirusgroup and are most closely related to another IncP-plasmid-dependent ssRNA phage, PRR1. Furthermore, our result strongly suggests that the stereotypical bauplans of genome organization found inLevivirusandAlloleviviruspredate phage specialization for conjugative plasmids, suggesting that the utilization of conjugative plasmids for cell attachment and entry comprises independent evolutionary events for these two main clades of ssRNA phages. Our result is also consistent with findings of a previous study, making theLevivirus-like genome organization ancestral and theAllolevivirus-like genome derived. To obtain a deeper insight into the evolution of ssRNA phages, more phages specializing for various conjugative plasmids and infecting different bacterial species would be needed.

Sign in / Sign up

Export Citation Format

Share Document