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Antigenic characterization of bovine ephemeral fever rhabdovirus G and GNS glycoproteins expressed from recombinant baculoviruses

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Abstract

Recombinant baculoviruses expressing the BEFV envelope glycoprotein G and non-structural glycoprotein GNS were constructed. The G protein expressed in insect cells was located on the cell surface and induced spontaneous cell fusion at mildly acidic pH. The expressed G protein reacted with MAbs to continuous and conformational neutralization sites (G1, G2, G3b and G4), but not to conformational site G3a. The expressed GNS protein was also located on the cell surface but did not exhibit fusogenic activity. The GNS protein reacted with polyclonal antiserum produced from vaccinia-virus-expressed recombinant GNS but did not react with G protein antibodies. A His6-tagged, soluble form of the G protein was expressed and purified by Ni2+–NTA chromatography. The purified G protein reacted with BEFV-neutralizing MAbs to all continuous and conformational antigenic sites. The highly protective characteristics of the native BEFV G protein suggest that the secreted, baculovirus-expressed product may be a useful vaccine antigen.

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Acknowledgments

We thank Lee Cadogan and Roger Pearson for their advice on the growth of High 5TM cells and the Ni2+–NTA chromatography, and Dr. Jeff Cowley and Dr. David Boyle for critical review of the manuscript. The plasmid pEXPORTHIS has been generously provided by Dr. Rosanne Casu. Dr. Jasjit Johal was supported by a CSIRO Postgraduate Research Scholarship.

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Author notes

  1. Jasjit Johal

    Present address: School of Health Sciences, University of Canberra, Kirinari Street, Bruce, Australia

  2. Karryn Gresty

    Present address: Malaria Drug Resistance and Chemotherapy Laboratory, Queensland Institute of Medical Research, and Drug Resistance and Diagnostics Australian Army Malaria Institute, Gallipoli Barracks, Enoggera, Australia

Authors and Affiliations

  1. CSIRO Livestock Industries, Queensland Bioscience Precinct, 308 Carmody Road, St Lucia, Qld, 4072, Australia

    Jasjit Johal, Karryn Gresty, Kritaya Kongsuwan & Peter J. Walker

  2. CSIRO Livestock Industries, Australian Animal Health Laboratory, 5 Portarlington Road, Geelong, Vic, 3220, Australia

    Peter J. Walker

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  1. Jasjit Johal
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Correspondence to Peter J. Walker.

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Johal, J., Gresty, K., Kongsuwan, K. et al. Antigenic characterization of bovine ephemeral fever rhabdovirus G and GNS glycoproteins expressed from recombinant baculoviruses. Arch Virol 153, 1657–1665 (2008). https://doi.org/10.1007/s00705-008-0164-0

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  • DOI: https://doi.org/10.1007/s00705-008-0164-0

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