Construction and characterization of a H19 epitope point mutant of MDV CVI988/rispens strain

Z. Cui, A. Qin, L. F. Lee, P. Wu, H. J. Kung

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Abstract

A recombinant virus, CVI/rpp38, was developed from the Marek's disease virus (MDV) CVI988/Rispens vaccine strain. This recombinant was obtained by transfection of CVI988/Rispens-infected chich embryo fibroblasts (CEFs) with plasmid pHA25 DNA containing pp38 gene from GA strain of MDV. Monoclonal antibody (MAb) H 19 which reacts with pp38 from GA but not with that from CV1998 was used to screen for recombinant viruses in transfected cell culture plates by immunofluorescent assay (IFA). A positive plaque was isolated, propagated, and purified from cell-free virus particles after sonication of infected CEFs. The mutant CVI/rpp38 was not only reactive with MAb H19 in IFA but also in immunoprecipitation. A 38 kDa protein was immunoprecipitated from the CVI/rpp38 mutant virus but not from parental CV1988 virus. DNA sequence of the mutant virus showed a substitution of G at position 320 by A resulting in a change of an amino acid residue from arginine to glutamine. Comparison of nucleotide sequence of pp38 from strains GA, Md5 and Md11/75c/R2 and CV1988 revealed change to glutamine in this position. The result of this study provides a direct evidence for the location of the identified HI 9 epitope in pp38. This mutant is potentially useful to further explore the biological function of pp38 and its H19 epitope.

Original languageEnglish
Pages (from-to)169-173
Number of pages5
JournalActa Virologica
Volume43
Issue number2-3
Publication statusPublished - Apr 1 1999
Externally publishedYes

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Keywords

  • MDV
  • Monoclonal antibody H19
  • Point mutant
  • pp38

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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