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.

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.

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.

INVESTIGATION ON BOVINE VIRAL DIARRHEA VIRUS AND INFECTIOUS BOVINE RHINOTRACHEITIS VIRUS IN MONGOLIAN DAIRY FARMS

2017 ◽  
Vol 19 (3) ◽  
pp. 10-15
Author(s):  
Dulam Purevtseren ◽  
Erdenechimeg Dashzevge ◽  
Zhou Wei Guan Guan
Keyword(s):  
Dairy Cows ◽  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Infectious Bovine Rhinotracheitis ◽  
Serum Samples ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
One Step ◽  
Viral Diarrhea Virus ◽  
Infectious Bovine Rhinotracheitis Virus

In this study, 168 blood sera were collected from dairy cows in Selenge and Tuv aimags during 2013 - 2014. The ELISA was carried out for serological detection of antibodies and antigens to Bovine viral diarrhea virus (BVDV), antibodies to Infectious bovine rhinotracheitis virus (IBRV) in dairy cows in Mongolia. The ELISA results of antibodies of BVDV and antigen of BVDV showed that 86.9% and 3.57%, respectively. The seroprevalence of antibodies against IBRV was found to be 60.7%. In order to confirm of BVDV, One Step RT-PCR was performed in ELISA positive cattle serum samples using specific primer for BVDV. The results showed that 294 bp fragment was successfully amplified. Phylogenetic analysis revealed that the nucleotide sequences 5'UTR gene of the isolates belonged to the BVDV1 subtype. Four isolated virus samples were closely related to China, the another isolate was closely related to the Slovenia BVDV1 isolate.

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