Archives of Virology

, Volume 71, Issue 1, pp 43–55 | Cite as

The genome of orf virus: Restriction endonuclease analysis of viral DNA isolated from lesions of orf in sheep

  • A. J. Robinson
  • G. Ellis
  • T. Balassu
Original Papers


The purification of orf virus directly from scab material from clinical cases of orf in sheep and restriction endonuclease analysis of the viral DNA is described. Between 7×109 and 1.6×1011 virus particles, and 0.7 to 22.8 µg of viral DNA could be recovered from lg of scab material. Considerable heterogeneity was observed between different field isolates when restriction endonuclease digests of orf DNA were compared by gel electrophoresis. It was also shown, for two isolates, that these fragment patterns did not change after plaque purification and passage in cell culture. it is suggested that restriction endonuclease analysis of viral DNA offers a convenient method of identification of isolates of orf virus. The molecular weight of orf DNA was determined and found to be 88.8×106.


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  1. 1.
    Backus, R. C., Williams, R. C.: The use of spraying methods of volatile suspending media in the preparation of specimens for electron microscopy. J. appl. Phys.21, 11–15 (1950).Google Scholar
  2. 2.
    Buchman, T. G., Roizman, B., Adams, G., Stover, J. H.: Restriction endonuclease fingerprinting of Herpes simplex virus DNA: A novel epidemiological tool applied to a nosocomial outbreak. J. inf. Dis.138, 488–498 (1978).Google Scholar
  3. 3.
    Davidson, N., Szybalski, W.: Physical and chemical characteristics of Lambda DNA. In:Hershey, A. D. (ed.), The Bacteriophage Lambda, 45–82. New York: Cold Spring Harbor Laboratory 1971.Google Scholar
  4. 4.
    Davis, R. W., Simon, M., Davidson, N.: Electron Microscope heteroduplex methods for mapping regions of base sequence homology in nucleic aids. In:Grossman, L., Moldave, K. (eds.), Methods in Enzymology, Vol. 21, 413–428. New York: Academic Press 1971.Google Scholar
  5. 5.
    Esposito, J. J., Obijeski, J. F., Nakano, J. H.: Orthopoxvirus DNA: Strain differentiation by electrophoresis of restriction endonuclease fragmented virion DNA. Virology89, 53–66 (1978).Google Scholar
  6. 6.
    Ferris, R. D., Plowright, W.: Simplified methods for the production of monolayers of testis cells from domestic animals, and for the serial examination of monolayer cultures. J. Path. Bact.75, 313–318 (1958).Google Scholar
  7. 7.
    Gangemi, J. D., Sharpe, D. G.: Use of a restriction endonuclease in analysing the genomes from two different strains of vaccinia virus. J. Virol.20, 319–323 (1976).Google Scholar
  8. 8.
    Gissman, L., Pfister, H., Zurhausen, H.: Human papilloma virus (HPV): Characterization of four different isolates. Virology76, 569–580 (1977).Google Scholar
  9. 9.
    Horgan, E. S., Haseeb, M. A.: The immunological relationships of strains of contagious pustular dermatitis virus. J. comp. Pathol.57, 1–7 (1947).Google Scholar
  10. 10.
    Linnemann, C. S., Buchanan, T. G., Light, I. J., Ballard, J. L., Roizman, B.: Transmission of Herpes Simplex virus type 1 in a nursery for the newborn: Identification of viral isolates by DNA fingerprinting. Lanceti, 964–966 (1978).Google Scholar
  11. 11.
    Lonsdale, M.: A rapid technique for distinguishing Herpes Simplex type 1 from type 2 by restriction enzyme technology. Lanceti, 848–852 (1979).Google Scholar
  12. 12.
    Menna, A., Wittek, R., Backmann, P. A., Mayr, A., Wyler, R.: Physical characterization of a stomatitis papulosa virus genome: A cleavage map for the restriction endonuclease Hind III and EcoR1. Arch. Virol.59, 145–156 (1979).Google Scholar
  13. 13.
    Miller, J. H.: Experiments in Molecular Genetics, 331. New York: Cold Spring Harbor Laboratory 1972.Google Scholar
  14. 14.
    Muller, H. K., Wittek, R., Schaffner, W., Schumperli, D., Menna, A., Wyler, R.: Comparison of five poxvirus genomes by analysis with restriction endonucleases Hind III, Bam I, and EcoR1. J. gen. Virol.38, 135–147 (1977).Google Scholar
  15. 15.
    Murray, K., Murray, N. E.: Phage lambda receptor chromosomes for DNA fragments made with restriction endonuclease III ofHaemophilus influenzae and restriction endonuclease I ofEscherichia coli. J. mol. Biol.98, 551–564 (1975).Google Scholar
  16. 16.
    Nagington, J., Horne, R. W.: Morphological studies of orf and vaccinia viruses. Virology16, 248–260 (1962).Google Scholar
  17. 17.
    Nagington, J., Newton, A. A., Horne, R. W.: The structure of orf virus. Virology23, 461–472 (1964).Google Scholar
  18. 18.
    Precausta, P., Stellman, Ch.: Isolation and comparative study “in vitro” of five strains of contagious ecthyma of sheep. Zbl. Vet. Med.B20, 340–355 (1973).Google Scholar
  19. 19.
    Sabine, M., Robertson, G. R., Whalley, J. M.: Differentiation of Sub-Types of Equine Herpes virus I by restriction endonuclease analysis. Aust. vet. J.57, 148–149 (1981).Google Scholar
  20. 20.
    Sawhney, A. N.: Studies on the virus of ecthyma contagiosum, III. Multiplicity of virus strains. Izvest. Microbiol. Inst. Sofia18, 179–183 (1966).Google Scholar
  21. 21.
    Sawhney, A. N.: Studies on the virus of ecthyma contagiosum, IV. A comparative study of different strains of the virus in the tissue culture. Izvest. Microbiol. Inst. Sofia18, 185–189 (1966).Google Scholar
  22. 22.
    Sharp, P. A., Sugden, B., Sambrook, J.: Detection of two restriction endonuclease activities inHaemophilus parainfluenzae using analytic agarose ethidium bromide electrophoresis. Biochemistry12, 3055–3063 (1973).Google Scholar
  23. 23.
    Thomas, M., Davis, R. W.: Studies on the cleavage of bacteriophage lambda DNA with EcoR1 restriction endonuclease. J. mol. Biol.91, 315–328 (1975).Google Scholar
  24. 24.
    Westwood, J. C. N., Harris, W. J., Zwartouw, H. T., Titmuss, D. M., Appleyard, G. J.: Studies on the structure of vaccinia virus. J. gen. Microbiol.34, 67 to 78 (1964).Google Scholar
  25. 25.
    Wittek, R., Kuenzle, C. C., Wyler, R.: High G+C content in parapoxvirus DNA. J. gen. Virol.43, 231–234 (1979).Google Scholar
  26. 26.
    Wittek, R., Herlyn, R., Schumperli, D., Bachman, P. A., Mayr, A., Wyler, R.: Genetic and antigenic heterogeneity of different parapoxvirus strains. Intervirology13, 33–41 (1980).Google Scholar
  27. 27.
    Younghusband, H. B., Bellett, A. J. D.: Mature form of the deoxyribonucleic acid from chick embryo lethal orphan virus. J. Virol.8, 265–274 (1971).Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • A. J. Robinson
    • 1
  • G. Ellis
    • 1
  • T. Balassu
    • 1
  1. 1.Department of Veterinary Pathology and Public HealthMassey UniversityPalmerston NorthNew Zealand

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