Abstract
The VP2 gene DNA (1.4 kb in approximate) of a very virulent infectious bursal disease virus (vvIBDV) Chinju strain detected in Chinju, Korea was cloned into the bacmid, a baculovirus shuttle vector, through transposition of the gene from initially cloned pFastBacHTa plasmid, a baculovirus expression vector, and was subsequently expressed in Spodoptera frugiperda (Sf) cells. Biological properties of the expressed VP2 subunit protein were characterized to aid in the development of genetically engineered diagnostic reagents and vaccines against the vvIVDV. When the VP2 DNA-recombinant bacmid was transfected and propagated in the Sf cells, the cells showed no occlusion formation, which is a positive evidence for the insertion of the VP2 DNA into the polyhedrin gene of the bacmid, whereas the occlusions were observed in the cells infected by the Autographa californica nuclear polyhedrosis virus, a wild baculovirus. The expression of VP2 DNA was identified by strong positive reaction in fluorescent antibody test using chicken anti-IBDV serum. The VP2 protein was determined as a polypeptide band with M r of 48 kDa by the sodium dodecyl-polyacrylamide gel electrophoresis for the lysate of the Sf cells infected with the recombinant bacmid. The VP2 protein was successfully purified from the cell lysate by Ni-NTA affinity chromatography. The expressed VP2 subunit protein reacted specifically with chicken anti-IBDV serum in Western blotting.
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Burkhardt E. and Muller H., Arch Virol 94, 297-303, 1987.
Kibenge F.S.B., Dhillon A.S., and Russell R.G., J Gen Virol 69, 1757-1775, 1988.
Murphy F.A., Gibbs E.P.J., Horzinek M.C., and Studdert M.J., Veterinary Virology, 3rd edn., Academic Press, San Diego, 1999, pp. 405-409.
Azad A.A., Barrett S.A., and Fahey K.J., Virology 143, 35-44, 1985.
Dobos P., Hill B.J., Hallett R., Kells D.T.C., Becht H., and Teninges D., J Virol 32, 593-605, 1979.
Fahey K.J., O'Donnell I.J., and Azad A.A., J Gen Virol 66, 1479-1488, 1985.
Mundt E., Beyer J., and Muller H., J Gen Virol 76, 437-443, 1995.
Mundt E., Kollner B., and Kretzschmar D., J Virol 71, 5647-5651, 1997.
Spies U., Muller H., and Becht H., Nucleic Acids Res 17, 7982, 1989.
Heine H.G., Haritou M., Failla P., Fahey K., and Azad A.A., J Gen Virol 72, 1835-1843, 1991.
Kang C.Y., Adv Virus Res 35, 177-192, 1988.
Azad A.A., McKern N.M., Macreadie I.G., Failla P., Heine H.G., Chapman A.J., Ward C.W., and Fahey K.J., Vaccine 9, 715-722, 1991.
Macreadie I.G., Vaughan P.R., Chapman A.J., McKern N.M., Jagadish M.N., Heine H.G., Ward C.W., Fahey K.J., and Azad A.A., Vaccine 8, 549-552, 1990.
Bayliss C.D., Peters R.W., Cook J.K.A., Reece R.L., Howes K., Binns M.M., and Boursnell M.E.G., Arch Virol 120, 193-205, 1991.
Heine H.G. and Boyle D.B., Arch Virol 131, 277-292, 1993.
Heine H.G., Hyatt A.D., and Boyle D.B., Virus Res 32, 313-328, 1994.
Darteil R., Bublot M., Laplace E., Bouquet J.F., Audonnet J.C., and Riviere M., Virology 211, 481-490, 1995.
Tsukamoto K., Kojima C., Komori Y., Tanimura N., Masaji M., and Yamaguchi S., Virology 257, 352-362, 1999.
Dybing J.K. and Jackwood D.J., Avian Dis 42, 80-91, 1988.
Jackwood D.J., Henderson K.S., and Jackwood R.J. Clin Diagn Lab Immunol 3, 456-463, 1996.
Vakharia V.N., Snyder D.B., He J., Edwards G.H., Savage P.K., and Mengel-Whereat S.A., J Gen Virol 74, 1201-1206, 1993.
Wang M.Y., Bentley W.E., and Vakharia V., Biotechnol Bioeng 43, 349-356, 1994.
Kim T.K. and Yeo S.G., Virus Genes 26(1), 97-106, 2003.
Tacken M.G.J., Rottier P.J.M., Gielkens A.L.J., and Peeters B.P.H., J Gen Virol 81, 209-218, 2000.
Laemmli U.K., Nature 227, 680-685, 1970.
Summer M.D. and Smith G.E., A Manual of Methods for Baculovirus Vector and Insect Cell Culture Procedure, Texas Agricultural Experiment Station, College Station, TX, 1987.
Martinez-Torrecuadrada J.L., Lazaro B., Rodriguez J.F., and Casal J.I., Clin Diagn Lab Immunol 7, 645-651, 2000.
Phenix K.V., Wark K., Luke C.J., Skinner M.A., Smyth J.A., Mawhinner K.A., and Todd D., Vaccine 19, 3116-3123, 2001.
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Kim, TK., Yeo, SG. Expression of VP2 Gene Protein of Infectious Bursal Disease Virus Detected in Korea. Virus Genes 27, 75–81 (2003). https://doi.org/10.1023/A:1025128619976
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DOI: https://doi.org/10.1023/A:1025128619976