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Isolation, characterization and immunolocalization of a globin-like antigen from Ostertagia ostertagi adults

  • D. C. De Graaf (a1), P. Berghen (a1), L. Moens (a2), T. M. De Marez (a1), S. Raes (a1), M. L. Blaxter (a3) and J. Vercruysse (a1)...


Western blot analysis using an anti-globin rabbit serum Rb94 revealed a major band of 17 kDa in extracts of Ostertagia ostertagi adults and 4th-stage larvae. The adult stage globin-like antigen (OoAdGlb) was purified from total worm extracts by liquid chromatography. The protein has an estimated molecular mass of 36 kDa under non-reducing conditions, suggesting a dimeric structure containing 2 non-covalently linked 17 kDa monomers. Tryptic peptides were sequenced and showed strong similarities with the globins of free-living and parasitic nematodes. Immunolocalization revealed the presence of this globin-like antigen in the body wall musculature and/or the cuticle of O. ostertagi adults. An enzyme-linked immunosorbent assay based on the purified OoAdGlb showed no differences in response between calves infected by O. ostertagi and/or Cooperia oncophora and the negative controls.


Corresponding author

*Corresponding author: University of Gent, Faculty of Veterinary Medicine, Department of Parasitology, Salisburylaan 133, B-9820 Merelbeke, Belgium. Tel: + 32 9 264 7400. Fax: +32 9 264 74 96.


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Ansorge, W. (1985). Fast and sensitive detection of protein and DNA bands by treatment with potassium permanganate. Journal of Biochemical and Biophysical Methods 11, 1320.
Blaxter, M. L. (1993). Nemoglobins: divergent nematode globins. Parasitology Today 9, 353–60.
Blaxter, M. L., Ingram, L. & Tweedie, S. (1994 a). Sequence, expression and evolution of the globins of the parasitic nematode Nippostrongylus brasiliensis. Molecular and Biochemical Parasitology 68, 114.
Blaxter, M. L., Vanfleteren, J. R., Xia, J. & Moens, L. (1994 b). Structural characterization of an Ascaris myoglobin. Journal of Biological Chemistry 269, 30181–6.
Burger, H. J., Eckert, J., Chevalier, H. J., Rahman, M. S. A. & Konigsmann, G. (1968). Parasitic gastroenteritis in bovines. 2. Trials on the experimental immunization of calves using irradiated larvae of Ostertagia and Cooperia. Veterinär Medizinische Nachrichten 4, 312–32.
Canals, A. & Gasbarre, L. C. (1990). Ostertagia ostertagi: isolation and partial characterization of somatic and metabolic antigens. International Journal for Parasitology 20, 1047–54.
Cross, D. A. & Klesius, P. H. (1989). Soluble extracts from larval Ostertagia ostertagi modulating immune function. International Journal for Parasitology 19, 5761.
De Baere, I., Liu, L., Moens, L., Van Beeumen, J., Gielens, C., Richelle, J., Trotman, C., Finch, J., Gerstein, M. & Perutz, M. (1992). Polar zipper sequence in the high-affinity hemoglobin of Ascaris suum: amino acid sequence and structural interpretation. Proceedings of the National Academy of Sciences, USA 89, 4638–42.
De Cock, H., Knox, D. P., Claerebout, E. & De Graaf, D. C. (1993). Partial characterization of proteolytic enzymes in different developmental stages of Ostertagia ostertagi. Journal of Helminthology 67, 271–8.
De Graaf, D. C., Berghen, P., Hilderson, H., Claerebout, E. & Vercruysse, J. (1994). Identification and isolation of a 19·7 kDa Ostertagia ostertagi specific antigen and evaluation of its potential for immunodiagnosis. International Journal for Parasitology 24, 681–8.
De Graaf, D. C., Peelman, L. J., Claerebout, E., Hilderson, H., Schallig, H. D. F. H. & Vercruysse, J. (1995). Cloning and sequencing of an excretory/secretory antigen from Ostertagia ostertagi fourth-stage larvae containing multiple tandem repeats. Molecular and Biochemical Parasitology 72, 239–41.
Di Iorio, E. E. (1981). Preparation of derivatives of ferrous and ferric hemoglobin. In Methods in Enzymology, vol. 76 (ed. Antonini, E., Rossi-Bernardi, L. & Chiancone, E.), pp. 5772.London: Academic Press.
Dixon, B., Walker, B., Kimmins, W. & Pohajdak, B. (1991). Isolation and sequencing of a cDNA for an unusual hemoglobin from the parasitic nematode Pseudoterranova decipiens. Proceedings of the National Academy of Sciences, USA 88, 5655–9.
Drabkin, D. L. & Austin, J. H. (1935). Spectrophotometric studies. II. Preparations from washed blood cells, nitric oxide, hemoglobin and sulfhemoglobin. Journal of Biological Chemistry 112, 51.
Frenkel, M. J., Dopheide, T. A. A., Wagland, B. M. & Ward, C. W. (1992). The isolation, characterization and cloning of a globin-like, host-protective antigen from the excretory-secretory products of Trichostrongylus colubriformis. Molecular and Biochemical Parasitology 50, 2736.
Herlich, H. & Douvres, F. W. (1979). Gastrointestinal nematode immunization trials in cattle. American Journal of Veterinary Research 40, 1781.
Herlich, H. & Tromba, F. G. (1982). Vaccine trials with ultraviolet attenuated Ostertagia ostertagi in calves. Proceedings of the Helminthological Society of Washington 49, 71–5.
Hilderson, H., Berghen, P., De Graaf, D. C., Claerebout, E. & Vercruysse, J. (1995). Immunisation of calves with Ostertagia ostertagi fourth stage larval antigens failed to protect calves from infection. International Journal for Parasitology 25, 757–60.
Hilderson, H., De Graaf, D. C., Vercruysse, J. & Berghen, P. (1993). Characterisation of Ostertagia ostertagi antigens by different bovine immunoglobulin isotypes. Research in Veterinary Science 55, 203–8.
Keith, K. A., Duncan, M. C., Murray, M., Bairden, K. & Tait, A. (1990). Stage-specific cuticular proteins of Ostertagia circumcincta and Ostertagia ostertagi. International Journal for Parasitology 20, 1037–45.
Laemmli, U. K. (1970). Cleavage of structural proteins during assembly of the head of the bacteriophage T4. Nature, London 227, 681–5.
Mansour, M. M., Dixon, J. B., Clarkson, M. J., Carter, S. D., Rowan, T. G. & Hammet, N. C. (1990). Bovine immune recognition of Ostertagia ostertagi larval antigens. Veterinary Immunology and Immunopathology 24, 361–71.
Pearson, W. R. & Lipman, D. J. (1988). Improved tools for biological sequence comparison. Proceedings of the National Academy of Sciences, USA 85, 2444–8.
Pratt, D., Boisvenue, R. J. & Cox, G. N. (1992). Isolation of putative cysteine protease genes of Ostertagia ostertagi. Molecular and Biochemical Parasitology 56, 3948.
Sulston, J., Du, Z., Thomas, K., Wilson, R., Hillier, L., Staden, R., Halloran, N., Green, P., Thierry-Mieg, J., Qiu, L., Dear, S., Coulson, A., Craxton, M., Durbin, R., Berks, M., Metzstein, M., Hawkins, T., Ainscough, R. & Waterston, R. (1992). The C. elegans genome sequencing project: a beginning. Nature, London 356, 3741.
Towbin, H., Staehelin, T. & Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Proceedings of the National Academy of Sciences, USA 76, 4350–4.
Vinogradov, S. N. (1985). The structure of invertebrate extracellular hemoglobins. Comparative Biochemistry and Physiology, B 82, 115.
Williams, J. C., Roberts, E. D. & Todd, J. M. (1974). Ostertagia ostertagi: establishment of patent infections in calves by intravenous inoculation. International Journal for Parasitology 4, 5562.


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Isolation, characterization and immunolocalization of a globin-like antigen from Ostertagia ostertagi adults

  • D. C. De Graaf (a1), P. Berghen (a1), L. Moens (a2), T. M. De Marez (a1), S. Raes (a1), M. L. Blaxter (a3) and J. Vercruysse (a1)...


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