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PMID: 2552412 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The characterization of the EBV alkaline deoxyribonuclease cloned and expressed in E. coli.

Nucleic acids research ·Vol. 17 ·No. 19 ·1989-10-11 ·Pages 7609-22

Baylis SA, Purifoy DJ, Littler E

Abstract

Studies of nucleic acid homology suggest the BGLF5 open reading frame of Epstein-Barr virus (EBV) encodes an alkaline deoxyribonuclease (DNase) sharing some homology with that of herpes simplex virus. We report here the expression of the BGLF5 open reading frame in E. coli and the expression of high levels of a novel alkaline DNase activity in induced cells. This alkaline DNase has been purified to apparent homogeneity as a single protein species. This is the first report of the expression of a herpesvirus coded DNase in a prokaryotic system and of the purification of the EBV DNase to demonstrable purity. It has the biochemical characteristics of a typical herpesvirus alkaline exonuclease showing a high pH optimum, an absolute requirement for Mg2+ for activity and sensitivity to high salt concentrations and polyamines. The enzyme activity was neutralized by sera from patients with nasopharyngeal carcinoma and was reactive with these sera in Western blot analysis. Thus the prokaryotic expression system described here provides an economical and efficient source of the EBV DNase for biochemical and seroepidemiological analysis.

MeSH Terms
Blotting, Western Cloning, Molecular DNA, Recombinant/metabolism Deoxyribonucleases/biosynthesis,genetics,metabolism Enzyme Induction Escherichia coli/genetics Genes, Viral Herpesvirus 4, Human/enzymology,genetics Kinetics Molecular Weight Plasmids Recombinant Proteins/biosynthesis,metabolism Viral Structural Proteins/genetics
Chemicals
DNA, Recombinant Recombinant Proteins Viral Structural Proteins Deoxyribonucleases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Baylis S A
Department of Molecular Biology, Paterson Institute for Cancer Research, Christie Hospital and holt Radium Institute, Manchester, UK.
Purifoy D J
Littler E
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.19.7609
Published
1989-10-11
Pages
7609-22
Language
English
Region
England
NLM ID
0411011
PMCID
PMC334870
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/17.19.7609 PMC Full Text View journal details
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