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PMID: 7685403 Published · ppublish English Journal Article

Gene mapping and expression of two immunodominant Epstein-Barr virus capsid proteins.

Journal of virology ·Vol. 67 ·No. 7 ·1993-07-00 ·Pages 3908-16

van Grunsven WM, van Heerde EC, de Haard HJ, Spaan WJ, Middeldorp JM

Abstract

The genomic localization of two immunodominant genes encoding two proteins of the Epstein-Barr virus capsid antigen (VCA) complex, VCA-p18 and VCA-p40, has been identified. For that purpose, lambda gt11-based cDNA libraries were constructed from HH514.c16 cells induced for virus production. The libraries were screened with a monoclonal antibody, EBV.OT41A, directed against VCA-p40 or with affinity-purified human antibodies against VCA-p18. Sequencing of the inserts of positive plaques showed that VCA-p18 and VCA-p40 are encoded within open reading frames (ORFs) BFRF3 and BdRF1, respectively. Peptide scanning analysis of the predicted protein of ORF BdRF1 resulted in defining the epitope of monoclonal antibody EBV.OT41A at the C-terminal region. The dominant VCA-p18 reactivity of human sera can be completely inhibited by preadsorption with Escherichia coli-expressed BFRF3-beta-galactosidase. Serum of a rabbit immunized with BFRF3-beta galactosidase reacts with a VCA-specific protein of 18 kDa. In addition, BFRF3-beta-galactosidase affinity-purified antibodies react with VCA-p18 of virus-producing cells (HH514.c16). Complete inhibition of viral DNA polymerase activity by phosphonoacetic acid is associated with the absence of RNAs and protein products of both ORFs, indicating that VCA-p18 and VCA-p40 are true late antigens.

MeSH Terms
Amino Acid Sequence Antibodies, Monoclonal/immunology Antibodies, Viral/immunology Antigens, Viral/genetics,immunology Base Sequence Blotting, Western Capsid/genetics,immunology Capsid Proteins Cloning, Molecular Epitopes Escherichia coli Genes, Viral Herpesvirus 4, Human/genetics,immunology Molecular Sequence Data Oligodeoxyribonucleotides/chemistry RNA, Viral/genetics Viral Structural Proteins/genetics
Chemicals
Antibodies, Monoclonal Antibodies, Viral Antigens, Viral Capsid Proteins Epitopes Epstein-Barr viral capsid antigen Oligodeoxyribonucleotides RNA, Viral Viral Structural Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
van Grunsven W M
Biotechnological Research Unit, Organon Teknika, RM Boxtel, The Netherlands.
van Heerde E C
de Haard H J
Spaan W J
Middeldorp J M
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.67.7.3908-3916.1993
Published
1993-07-00
Pages
3908-16
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC237757
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.67.7.3908-3916.1993 PMC Full Text View journal details
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