1887

Abstract

The gene encoding the envelope glycoprotein of a recent Danish isolate of a salmonid rhabdovirus, viral haemorrhagic septicaemia virus (VHSV) has been cloned and sequenced at the cDNA level. When compared with the deduced sequence of a French isolate of VHSV, it was noted that there were 13 amino acid substitutions in the Danish virus. Amino acid homologies with the glycoprotein of a North American salmonid rhabdovirus (infectious haematopoietic necrosis virus) indicate a high degree of structural similarity between the two fish rhabdovirus glycoproteins. Results from partial enzymatic deglycosylation of the viral protein indicate that all four NXT/S sites found in the sequence are -glycosylated in the virus. The glycoprotein, without the N-terminal leader sequence and C-terminal hydrophobic anchor segment, was expressed in as a factor X protease-cleavable fusion protein. The purified and renatured viral part of the recombinant protein was able to elicit VHSV-specific antibodies and neutralizing antibody activity in serum when injected into rainbow trout.

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1993-04-01
2021-10-25
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References

  1. Celis E. 1990; Rhabdoviruses. In Immunochemistry of Viruses II. The Basis for Serodiagnosis and Vaccines pp 345–358 Edited by van Regenmortel M. HV., Neurath A. R. Amsterdam: Elsevier;
    [Google Scholar]
  2. Chomczynski P., Sacchi N. 1987; Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analytical Biochemistry 162:156–159
    [Google Scholar]
  3. De kinkelin P. 1988; Vaccination against viral haemorrhagic septicaemia. In Fish Vaccination pp 172–192 Edited by Ellis A. E. London: Academic Press;
    [Google Scholar]
  4. Dorson M., Torchy C. 1979; Complement dependent neutralization of Egtved virus by trout antibodies. Journal of Fish Diseases 2:345–347
    [Google Scholar]
  5. Elder H., Alexander S. 1982; Endo-β-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins. Proceedings of the National Academy of Sciences, U.S.A. 79:4540–4544
    [Google Scholar]
  6. Fryer J. L., Yusha A., Pilcher K. 1965; The in vitro cultivation of tissue and cells of Pacific salmon and steelhead trout. Annals of the New York Academy of Sciences 126:566–586
    [Google Scholar]
  7. Gilmore R. D. Jr, Engelking H. M., Scott manning D., Leong J. C. 1988; Expression in Escherichia coli of an epitope of the glycoprotein of infectious hematopoietic necrosis virus protects against viral challenge. Bio/Technology 6:295–300
    [Google Scholar]
  8. Hochuli E., Bannwart W., Döbeli H., Gentz R., Strüber D. 1988; Genetic approach to facilitate purification of recombinant proteins with a novel metal chelate adsorbent. Bio/Technology 6:1321–1325
    [Google Scholar]
  9. Jensen M. H. 1965; Research on the virus of Egtved disease. Annals of the New York Academy of Sciences 126:422–426
    [Google Scholar]
  10. Jørgensen P. EV., Olesen N. J., Lorenzen N., Winton J. R., Ristow S. S. 1991; Infectious hematopoietic necrosis (IHN) and viral hemorrhagic septicaemia (VHS): detection of trout antibodies to the causative viruses by means of plaque neutralization, immunofluorescence, and enzyme-linked immunosorbent assay. Journal of Aquatic Animal Health 3:100–108
    [Google Scholar]
  11. Kelley J. M., Emerson S., Wagne R. 1972; The glycoprotein of vesicular stomatis virus is the antigen that gives rise to and reacts with neutralizing antibody. Journal of Virology 10:1231–1235
    [Google Scholar]
  12. Koener J. F., Passavant C. W., Kurath G., Leong J. 1987; Nucleotide sequence of a cDNA clone carrying the glycoprotein gene of infectious hematopoietic necrosis virus, a fish rhabdovirus. Journal of Virology 61:1342–1349
    [Google Scholar]
  13. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  14. Lorenzen N., Olesen N., Jørgensen P. EV. 1988; Production and characterization of monoclonal antibodies to four Egtved virus structural proteins. Diseases of Aquatic Organisms 4:35–42
    [Google Scholar]
  15. Lorenzen N., Olesen N. J., Jørgensen P. EV. 1990; Neutralization of Egtved virus pathogenicity to cell cultures and fish by monoclonal antibodies to the viral G protein. Journal of General Virology 71:561–567
    [Google Scholar]
  16. Lorenzen N., Olesen N. J., Jørgensen P. EV. 1993; Antibody response to VHS virus proteins in rainbow trout. Journal of Fish and Shellfish Immunology (in press)
    [Google Scholar]
  17. Malek L., Soostmayer G., Garvin R. T., James E. 1984; The rabies glycoprotein gene is expressed in E. coli as a denatured polypeptide. In Modern Approaches to Vaccines pp 203–208 Edited by Chanock R. M., Lerner R. A. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  18. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  19. Matsumura M., Signor G., Matthews B. W. 1989; Substantial increase of protein stability by multiple disulphide bonds. Nature, London 342:291–293
    [Google Scholar]
  20. Mierendorf R. C., Percy C., Young R. A. 1987; Gene isolation by screening λgtl1 libraries with antibodies. Methods in Enzymology 152:458–469
    [Google Scholar]
  21. Nagai K., Thogersen H. C. 1987; Synthesis and sequence-specific proteolyses of hybrid proteins produced in Escherichia coli. Methods in Enzymology 153:461–481
    [Google Scholar]
  22. Olesen N. J., Jørgensen P. E. 1986; Detection of neutralizing antibody to Egtved virus in rainbow trout (Salmo gairdneri) by plaque neutralization test with complement addition. Journal of Applied Ichthyology 2:33–41
    [Google Scholar]
  23. Rose J., Schubert M. 1987; Rhabdovirus genomes and their products. In The Rhabdoviruses pp 129–166 Edited by Wagner R. R. New York: Plenum Press;
    [Google Scholar]
  24. Sakai R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. 1988; Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
    [Google Scholar]
  25. SJOBLOM I., LUNDSTROM M., SJOGREN-JANSSON E., GLORIOSO J. C., JEANSSON S. 1987; Demonstration and mapping of highly carbohydrate-dependent epitopes in the herpes simplex virus type 1-specified glycoprotein C. Journal of General Virology 68:545–554
    [Google Scholar]
  26. THIRY M. 1991; Viral haemorrhagic septicaemia virus M2 gene for matrix protein (C-term.) and gene for glycoprotein (envelope spike protein). EMBL/GenBank Data Libraries, accession no. X59148.
    [Google Scholar]
  27. Thiry M., Lecocq-xhonneux F., Dheur I., Renard A., De kiykelin P. 1991; Sequence of a cDNA carrying the glycoprotein gene and part of the matrix protein M2 gene of viral haemorrhagic septicaemia, a fish rhabdovirus. Biochimica et biophysica acta 1090:345–347
    [Google Scholar]
  28. Thuvander A., Fossum C., Lorenzen N. 1990; Monoclonal antibodies to salmonid immunoglobulin: characterization and applicability in immunoassays. Developmental and Comparative Immunology 14:415–423
    [Google Scholar]
  29. Vidal S., Mottet G., Kolakofsky D., Roux L. 1989; Addition of high-mannose sugars must precede disulfide bond formation for proper folding of Sendai virus glycoproteins. Journal of Virology 63:892–900
    [Google Scholar]
  30. Wolf K., Gravell M., Malsberger R. 1966; Lymphocystis virus: isolation and propagation in centrarchid fish cell lines. Science 151:1004–1005
    [Google Scholar]
  31. Xu L., Mourich D., Engelking H. M., Ristow S., Arnzen J., Leone J. 1991; Epitope mapping and characterization of the infectious hematopoietic necrosis virus glycoprotein, using fusion proteins synthesized in Escherichia coli. Journal of Virology 65:1611–1615
    [Google Scholar]
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