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- Title
Bacterial Synthesis of Herpes Simplex Virus Types 1 and 2 Glycoprotein D Antigens.
- Authors
Watson, Roger J.; Weis, John H.; Salstrom, John S.; Enquist, Lynn W.
- Abstract
We have used elements of the <em>E. coli</em> lactose (<em>lac</em>) operon to produce a collection of herpes simplex virus types 1 and 2 glycoprotein D (gD-1 and gD-2) antigens. Our approach employed recombinant DNA techniques to construct plasmids with various segments of the gD-1 and gD-2 coding sequences fused to the <em>lac</em>Z gene. Such hybrid genes were expressed in a regulated manner in <em>E. coli</em> by joining them to the lac promoter-operator region. Efficient translation of these hybrid genes was facilitated by incorporating a coding sequence specifying a short peptide leader (λ<em>cro</em>) in the plasmid expression vectors resulting in synthesis of chimeric Cro-gD-β-galactosidase proteins. In addition, insertion of synthetic translation terminators at the junction of gD and <em>lac</em>Z enabled us to produce specific truncated gD polypeptide sequences unfused to β-galactosidase. The gD antigens produced in <em>E. coli</em> were not glycosylated and were generally recovered as dense insoluble aggregates. Proteins containing portions of gD- 1 or gD- 2 were analyzed by immunoprecipitation using anti-HSV rabbit serum and a number of monoclonal antibodies recognizing different epitopes of gD- 1. Initial animal studies were done with antigens that reacted with neutralizing antisera or monoclonal antibodies. When these bacterially produced proteins were injected into rabbits, antibodies were produced that specifically immunoprecipitated authentic gD polypeptides and neutralized the infectivity of both virus types. These studies suggest that gene fusion techniques can be used to produce immunogenic proteins in large quantity. These polypeptides are not only useful in analyses of gene structure and function, but also can provide novel diagnostic reagents and well-defined pure antigens for vaccine development.
- Subjects
HERPES simplex virus; MICROBIOLOGICAL synthesis; GLYCOPROTEINS; ANTIGENS; ESCHERICHIA coli; DNA; IMMUNOGLOBULINS
- Publication
Journal of Investigative Dermatology, 1984, Vol 83, p102s
- ISSN
0022-202X
- Publication type
Article
- DOI
10.1038/jid.1984.30