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    Comparison of thymidine kinase and A‐type inclusion protein gene sequences from Norwegian and Swedish cowpox virus isolates
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    Abstract:
    During the last decades, cowpox virus, a member of the genus Orthopoxvirus within the Poxviridae family, has appeared as a pathogen in domestic cats, zoo animal species, and humans. At the same time, vaccinia virus, another orthopoxvirus, has been used as a recombinant vaccine vector with foreign genes inserted in the thymidine kinase (TK) gene. By PCR and cycle sequencing, we have determined the nucleotide sequences of the TK gene and the A‐type inclusion protein (ATIP) gene of virus isolates from two human cowpox cases in Sweden, as well as a human and a feline case from Norway. We also obtained the corresponding sequences from ectromelia virus (strain Moscow), cow‐pox virus (strain Brighton) and vaccinia virus (strain Western Reserve). The new virus isolates differed from ectromelia virus and vaccinia virus, and were confirmed to be cowpox virus strains. Isolates originating from the same country had nearly identical TK sequences and fully identical ATIP sequences. They probably represent local geographical strains of cowpox virus.
    Keywords:
    Cowpox virus
    Ectromelia virus
    Orthopoxvirus
    Cowpox
    Poxviridae
    Recombinant virus
    Fowlpox virus
    During the last decades, cowpox virus, a member of the genus Orthopoxvirus within the Poxviridae family, has appeared as a pathogen in domestic cats, zoo animal species, and humans. At the same time, vaccinia virus, another orthopoxvirus, has been used as a recombinant vaccine vector with foreign genes inserted in the thymidine kinase (TK) gene. By PCR and cycle sequencing, we have determined the nucleotide sequences of the TK gene and the A‐type inclusion protein (ATIP) gene of virus isolates from two human cowpox cases in Sweden, as well as a human and a feline case from Norway. We also obtained the corresponding sequences from ectromelia virus (strain Moscow), cow‐pox virus (strain Brighton) and vaccinia virus (strain Western Reserve). The new virus isolates differed from ectromelia virus and vaccinia virus, and were confirmed to be cowpox virus strains. Isolates originating from the same country had nearly identical TK sequences and fully identical ATIP sequences. They probably represent local geographical strains of cowpox virus.
    Cowpox virus
    Ectromelia virus
    Orthopoxvirus
    Cowpox
    Poxviridae
    Recombinant virus
    Fowlpox virus
    We have sequenced DNA equivalent to the E5R ORF of Copenhagen vaccinia virus from an additional strain of vaccinia and from cowpox (three strains), camelpox (two strains), taterapox and ectromelia viruses. None of these showed the disruptions previously reported in the equivalent region of monkeypox virus. We also constructed a viable recombinant of vaccinia virus strain Dairen in which the E5R sequence was disrupted by a 436 bp deletion and substitution of the E. coli gpt gene. Quantitative analysis of the sequences, including available sequences from monkeypox, variola and vaccinia viruses revealed four main groupings, namely cowpox, ectromelia, monkeypox and a cluster which includes variola, camelpox, taterapox and vaccinia viruses. It was noted that, at over 75% of the positions which differentiated species, all species but one had a common nucleotide. Although the analysis covers one single gene only, the results accord with what is known of the biology of the viruses.
    Cowpox
    Monkeypox
    Cowpox virus
    Orthopoxvirus
    Poxviridae
    Ectromelia virus
    Ectromelia
    Strain (injury)
    Citations (17)
    Ectromelia virus
    Cowpox virus
    Orthopoxvirus
    Cowpox
    Affinities
    Poxviridae
    Dissociation constant
    Citations (70)
    SUMMARY Orthopoxvirus DNA from representative strains of rabbitpox, vaccinia, monkeypox, variola, cowpox and ectromelia viruses was analysed by cleavage with restriction endonucleases HindIII, Xho1 or Sma1. Genome mol. wt. vary from about 120 × 106 for rabbitpox to about 145 × 106 for cowpox. Physical maps of cleavage sites are similar and characteristic for strains of the same Orthopoxvirus type. The distribution of HindIII sites suggests that an internal region of mol. wt. about 30 × 106 is highly conserved between Orthopoxvirus genomes although some type-specific differences occur within this region, especially with strains of ectromelia virus. Conservation of internal sequences is less marked following analysis with Xho1 although cleavages within this central region of particular genomes appear to represent a subset of preferred sites. Endonuclease Sma1 cleaves exceptionally infrequently and distinguishes variola, monkeypox, vaccinia, cowpox or ectromelia viruses. Type specific differences result largely from extensive, near terminal variations in length and sequence. Representative Orthopoxvirus genomes have rapidly renaturing terminal restriction fragments confirming the presence of near terminal, covalent cross-links. Terminal restriction fragments from the same or different genomes generally cross hybridize indicating the presence of near terminal repetitions of mol. wt. up to 6 × 106 and which share at least a subset of common sequences. Variola strains however, appear to lack such sequences from one specific terminus which maps shorter than that of related viruses.
    Orthopoxvirus
    Ectromelia virus
    Monkeypox
    Cowpox
    Cowpox virus
    Variola virus
    Ectromelia
    Poxviridae
    Citations (228)
    2 pox outbreaks among white rats in a breeding colony are described. The infection occurred in 3 different forms: pulmonary, dermal and mixed. Apparently healthy animals appeared to be virus carriers. The virus isolated belonged to the genus Orthopoxvirus of Poxviridae family, and was very close to cowpox virus. It differed from reference strains of cowpox virus in having a lower ceiling temperature and a higher pathogenicity for white rats.
    Cowpox virus
    Orthopoxvirus
    Poxviridae
    Cowpox
    Pox virus
    Ectromelia virus
    Fowlpox virus
    Variola virus
    White (mutation)
    Citations (23)
    Concern regarding the use of variola and monkeypox viruses as bioterrorist agents has led to an increased study of orthopoxviruses to understand the molecular and cellular basis of pathogenesis and develop safe and effective antivirals and vaccines against smallpox. Crucial to these efforts is the availability of animal models, which are inexpensive, genetically homogeneous, and recapitulate the human disease. The popular small-animal orthopoxvirus models employ the inbred mouse as the host, the respiratory tract as the site of virus inoculation, and orthopoxviruses—vaccinia, cowpox, and ectromelia viruses—as surrogates for variola virus. Ectromelia virus is likely the best surrogate for variola virus in a mouse model, as it is infectious at very low doses of virus, and the mousepox disease is associated with high mortality in the susceptible A, BALB/c, and DBA/2 stains of mice, but causes an unapparent infection in the C57BL/6 mouse strain. This chapter describes an ectromelia virus respiratory infection model in the mouse.
    Ectromelia virus
    Monkeypox
    Orthopoxvirus
    Cowpox
    Variola virus
    Ectromelia
    Poxviridae
    Cowpox virus
    Citations (29)
    Ectromelia virus
    Orthopoxvirus
    Cowpox
    Variola virus
    Cowpox virus
    Ectromelia
    Poxviridae