Expression of an early Epstein-Barr virus antigen (EA-D) in E. coli

Roeckel, Dagmar; Boos, Harald; Mueller-Lantzsch, N.

doi:10.1007/bf01314434

Cited by 4 publications

References 29 publications

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Immunological characterization of the epstein‐barr virus phosphoprotein pp58 and deoxyribonuclease expressed in the baculovirus expression system

Chen¹,

Sauter²,

Haiss³

et al. 1991

Intl Journal of Cancer

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The open reading frames of the phosphoprotein pp58 (BMRFI) and the deoxyribonuclease (BGLF5) of the Epstein-Barr-virus (EBV) strain M-ABA were cloned in the baculovirus expression vectors pAc373 and pAc360 and expressed in the Spodoptera frugiperda (SF158) insect cells. The recombinant phosphoprotein pp58 expressed in SF158 cells was recognized by the anti-pp58 rabbit anti-sera which were generated by immunizing rabbits with a TrpE-BMRFI fusion protein expressed in E. coli. DNA-cellulose chromatography showed that the recombinant pp58 exhibited DNA-binding activities. Immunofluorescence, immunoblot and ELISA analysis indicated that sera from patients with nasopharyngeal carcinoma (NPC) contained antibodies against pp58. The recombinant EBV DNase expressed in SF158 cells was recognized by the anti-EBV DNase rabbit anti-sera which were generated by immunizing rabbits with a TrpE-C-terminal part of BGLF5 fusion protein expressed in E. coli. The anti-EBV DNase rabbit anti-sera recognized also a protein of about 52 kDa in the EBV-harboring human B-cell lines Raji, Jijoye, B95-8, M-ABA and BL74 induced by TPA and n-butyrate. The recombinant EBV DNase exhibited exonuclease and endonuclease activities, a requirement for magnesium, and a high pH optimum (8.0). Its enzyme activities could be inhibited by sera from NPC patients and anti-EBV DNase rabbit anti-sera. Comparable studies of Raji EBV-DNase and recombinant EBV-DNase implied that recombinant EBV-DNase could also be used in the enzyme activity assay for the detection of NPC. In contrast to the enzyme inhibition test, immunofluorescence and immunoblot analysis demonstrated that the recombinant EBV DNase exhibited only a weak immunological reaction with NPC sera.

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Immunological characterization of the epstein‐barr virus phosphoprotein pp58 and deoxyribonuclease expressed in the baculovirus expression system

Chen¹,

Sauter²,

Haiss³

et al. 1991

Intl Journal of Cancer

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EBV PP58 and DNase Expressed in the Baculovirus System May Improve the Diagnosis of NPC

Chen

Sauter

Heiss

et al. 1991

Epstein-Barr Virus and Human Disease • 1990

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Mapping of viral epitopes with prokaryotic expression products

Lenstra

Kusters

Zeijst

1990

Archives of Virology

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Several systems are available for the expression of foreign gene sequences in Escherichia coli. We describe the use of prokaryotic expression products of viral gene fragments in order to identify the regions that specify the binding sites of antibodies. This approach is particularly successful if the antigenicity does not depend on the native protein, but only on the amino acid sequence, i.e., if the epitope is sequential. Combining prokaryotic expression with the use of synthetic peptides often permits a fast and accurate mapping of an epitope. The occurrence of immunodominant sequential epitopes on the surfaces of viruses seems to be a widespread phenomenon.

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Expression of an early Epstein-Barr virus antigen (EA-D) in E. coli

Cited by 4 publications

References 29 publications

Immunological characterization of the epstein‐barr virus phosphoprotein pp58 and deoxyribonuclease expressed in the baculovirus expression system

Immunological characterization of the epstein‐barr virus phosphoprotein pp58 and deoxyribonuclease expressed in the baculovirus expression system

EBV PP58 and DNase Expressed in the Baculovirus System May Improve the Diagnosis of NPC

Mapping of viral epitopes with prokaryotic expression products

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