Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
|
| pubmed:dateCreated |
1997-3-4
|
| pubmed:databankReference | |
| pubmed:abstractText |
A glycoprotein B (gB) gene homologue was identified in a 5.4-kb BamHl genomic fragment of the phocid herpesvirus type-1 (PhHV-1) which represents a widespread and important pathogen of pinnipeds. Sequence analysis revealed a gB-specific open-reading frame comprising 881 amino acids. Phylogenetic analysis gave evidence for a close evolutionary relationship between PhHV-1 and members of the Varicellovirus genus of the alpha-Herpesvirinae and canid herpesvirus in particular. In PhHV-1-infected Crandell feline kidney cells gB is expressed as a 113-kDa glycosylated molecule which is proteolytically cleaved into at least two fragments of 67 and 53-59 kDa apparently forming disulfide-linked heterodimers of 140 kDa. Cell surface expression of PhHV-1 gB was confirmed by FACS analysis. Thus, synthesis and processing of the gB protein of PhHV-1 follows a pattern also observed in other Varicelloviruses. Since the gB protein of herpesviruses, expressed in the baculovirus system, has been shown to be a suitable target for vaccine design, we used this system for expression of PhHV-1 gB. Recombinant (rec) baculovirus-expressed gB was identified as a 105-kDa glycosylated molecule. Proteolytic cleavage into fragments of 62 and 52 kDa was markedly delayed compared to wild-type (wt) gB. Wt and rec gB harbored endoglycosidase H (precursor)- as well as N-glycosidase F-sensitive N-glycans (proteolytic fragments). Baculovirus-expressed gB appeared to be antigenically authentic, since it was recognized in radioimmunoprecipitation and immune peroxidase monolayer assays by PhHV-1-neutralizing seal sera and by gB-specific neutralizing murine monoclonal antibodies. Furthermore, PhHV-1-neutralizing antibodies were induced in mice following immunization with baculovirus-expressed gB, indicating its suitability for incorporation in a candidate vaccine for seals.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Jan
|
| pubmed:issn |
0042-6822
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
20
|
| pubmed:volume |
227
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
343-52
|
| pubmed:dateRevised |
2006-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:9018133-Amino Acid Sequence,
pubmed-meshheading:9018133-Animals,
pubmed-meshheading:9018133-Baculoviridae,
pubmed-meshheading:9018133-Base Sequence,
pubmed-meshheading:9018133-Cats,
pubmed-meshheading:9018133-Cell Line,
pubmed-meshheading:9018133-Cloning, Molecular,
pubmed-meshheading:9018133-DNA, Viral,
pubmed-meshheading:9018133-Genes, Viral,
pubmed-meshheading:9018133-Kidney,
pubmed-meshheading:9018133-Mice,
pubmed-meshheading:9018133-Molecular Sequence Data,
pubmed-meshheading:9018133-Open Reading Frames,
pubmed-meshheading:9018133-Phylogeny,
pubmed-meshheading:9018133-Polymerase Chain Reaction,
pubmed-meshheading:9018133-Recombinant Proteins,
pubmed-meshheading:9018133-Restriction Mapping,
pubmed-meshheading:9018133-Seals, Earless,
pubmed-meshheading:9018133-Spodoptera,
pubmed-meshheading:9018133-Transfection,
pubmed-meshheading:9018133-Varicellovirus,
pubmed-meshheading:9018133-Viral Envelope Proteins,
pubmed-meshheading:9018133-Viral Structural Proteins
|
| pubmed:year |
1997
|
| pubmed:articleTitle |
Molecular characterization and baculovirus expression of the glycoprotein B of a seal herpesvirus (phocid herpesvirus-1).
|
| pubmed:affiliation |
Department of Virology, Erasmus University Rotterdam, The Netherlands.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|