Abstract
Marek's disease virus (MDV) gene clones, RA2 and GA8, constructed inE. coli bacteriophage lambda-gt11 (gt11) were identified by a monoclonal antibody (MAb), H19.47, against a putative transformation-related viral antigen consisting of a complex of three phosphorylated polypeptides, pp41, pp38, and pp24. Both recombinants have a MDV-DNA insert of about 0.5 kb and are mapped to the region ofBamHI-H orEcoRI-X fragments of the MDV genome by Southern blot hybridization. Immunoblot and immunoprecipitation with H19.47 identified a recombinant beta-galactosidase-MDV 140-kD fusion protein for RA2 and a 127-kD fusion protein for GA8.
Immunoprecipitation of35S-methionine-labeled, MDV-infected chicken embryo fibroblasts (CEF) with antisera against RA2 and GA8 fusion proteins recognized five polypeptides, of which three (p41, p38, and p24) are specified by H19.47 and the remaining two, p135 and p20, have not been previously identified. Immunoprecipitation of32P-phosphate-labeled or3H-glucosamine-labeled, GA-MDV-infected CEF with the antiserum against RA2 fusion protein identified a phosphorylated polypeptide of 38 kD and two glycoproteins of 60 and 49 kD, respectively. The antisera against recombinant fusion proteins thus revealed the existence of epitopes common to the phosphorylated polypeptides and other MDV-specific polypeptides.
Sera from chickens or mice hyperimmunized with the purified fusion proteins reacted with serotype 1, MDV-infected CEF in the fluorescent antibody (FA) test to significant titers. These immune sera did not react with either serotype II or III, indicating the serotype specificity of the phosphorylated polypeptides.
Article PDF
Similar content being viewed by others
References
Witter R. L., Nazerian K., Purchase H. G. and Burgoyne G. H., Am J Vet Res31, 525–538, 1970.
Okazaki W., Purchase H. G. and Burmester B. R., Avian Dis14, 413–429, 1970.
Churchill A. E., Chubb R. C. and Baxendale W., J Gen Virol4, 557–564, 1969.
Isfort R. J., Kung H-J. and Velicer L. F., J Virol61, 2614–2620, 1987.
Glaubiger C., Nazerian K. and Velicer L. F., J Virol45, 1228–1234, 1983.
Long P. A., Kaveh-Yamini P. and Velicer L. F., J Virol15, 1182–1191, 1975.
Long P. A., Clark J. L. and Velicer L. F., J Virol15, 1192–1201, 1975.
Isfort R. J., Sithole I., Kung H.J. and Velicer L. F., J Virol59, 411–419, 1986.
Silva R. F. and Lee L. F., Virology136, 307–320, 1984.
Ikuta K., Nishi Y., Kato S. and Hirai K., Virology114, 277–281, 1981.
Zaane D. V., Brinkhof J. M. A., Westenbrink F. and Gielkens A. L. J., Virology121, 116–132, 1982.
Lee L. F., Liu X. and Witter R. L., J Immunol130, 1003–1006, 1983.
Fukuchi K., Sudo M., Lee Y-S., Tanaka A., and Nonoyama M., J Virol51, 102–109, 1984.
Gibbs C. P., Nazerian K., Velicer L. F. and Kung H-J., Proc Natl Acad Sci USA81, 3365–3369, 1984.
Igarashi T., Takahashi M., Donovan J., Jessip J., Smith M., Hirai K., Tanaka A. and Nonoyama M., Virology157, 351–358, 1987.
Silva R. F. and Witter R. L., J Virol54, 690–696, 1985.
Buckmaster A., Scott S. D., Sanderson M. J., Boursnell M. E. G., Ross N. L. J. and Binns M. M., J Gen Virol69, 2033–2042, 1988.
Ross L. J. N., Sanderson M., Scott S. D., Binns M. M., Doel T. and Milne B., J Gen Virol70, 1789–1804, 1989.
Coussens P. M. and Velicer L. F., J Virol62, 2373–2379, 1988.
Bradley G., Hayashi M., Lancz G., Tanaka A. and Nonoyama M., J Virol63, 2534–2542, 1989.
Sithole I., Coussens P. M., Lee L. F., and Velicer L. F. in Kato S., Horiuchi T., Mikami T. and Hirai K. (eds.)Advances in Marek's Disease Research. Osaka, Japan, 1988, pp. 148–154.
Ikuta K., Nakajima K., Naito M., Ann S. H., Ueda S., Kato S. and Hirai K., Int J Cancer35, 257–264, 1985.
Nakajima K., Ikuta K., Naito M., Ueda S., Kato S. and Hirai K., J Gen Virol68, 1379–1389, 1987.
Radke K. and Martin G. S., Proc Natl Acad Sci USA76, 5212–5216, 1979.
Young R. A. and Davis R. W., Proc Natl Acad Sci USA80, 1194–1198, 1983.
Mocarski E. S., Pereira L. and Michael N., Proc Natl Acad Sci USA82, 1266–1270, 1985.
Petrovskis E. A., Timmins J. G. and Post L. E., J Virol60, 185–193, 1986.
Allen G. P. and Yeargan M. R., J Virol61, 2454–2461, 1987.
Banapour B., Rosenthal K., Rabin L., Sharma V., Young L., Fernandez J., Engleman E., McGrath M., Reyes G. and Lifson J., J Immunol139, 4027–4033, 1987.
Rodriguez J. F. and Esteban M., J Virol61, 3550–3554, 1987.
Eidson C. S. and Schmittle S. C., Avian Dis12, 467–476, 1968.
Purchase H. G., J Virol3, 557–565, 1969.
Witter R. L., Silva R. F., and Lee L. F., Avian Dis31, 829–840, 1987.
Maniatis T., Fritsch E. F. and Sambrook J.,Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1982.
Witter R. L., Purchase H. G. and Burgoyne G. H., J Natl Cancer Inst45, 567–577, 1970.
Author information
Authors and Affiliations
Additional information
Requests for reprints should be addressed to Lucy F. Lee, USDA, Agricultural Research Service, Regional Research Laboratory, 3606 East Mount Hope Road, East Lansing, MI 48823, USA.
Rights and permissions
About this article
Cite this article
Cui, Z., Yan, D. & Lee, L.F. Marek's disease virus gene clones encoding virus-specific phosphorylated polypeptides and serological characterization of fusion proteins. Virus Genes 3, 309–322 (1990). https://doi.org/10.1007/BF00569038
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00569038