alexandersen-productionofminkenteritis-1994.pdf (2.08 MB)
Production of mink enteritis parvovirus empty capsids by expression in a baculovirus vector system: a recombinant vaccine for mink enteritis parvovirus in mink
journal contribution
posted on 1994-01-01, 00:00 authored by J Christensen, Soren AlexandersenSoren Alexandersen, B Bloch, B Aasted, A UttenthalThe VP-2 gene of mink enteritis parvovirus (MEV) was amplified by the polymerase chain reaction using MEV DNA isolated from the faeces of a naturally infected mink. Subsequently the VP-2 gene was cloned into a baculovirus expression vector. Recombinant baculoviruses were isolated and the MEV VP-2 gene product was characterized after expression in Sf9 insect cells. The MEV VP-2 product had the same size as that reported for the wild-type MEV VP-2 protein and was recognized by convalescent sera from MEV-infected mink and a panel of monoclonal antibodies reactive to MEV. Furthermore, the VP-2 protein was able to form parvovirus-like particles, which had haemagglutinating properties comparable with the wild-type MEV. The cloned VP-2 gene was sequenced and only five nucleotide differences were found after alignment with the known sequences of the MEV type 1 and type 2 isolates. Surprisingly, the VP-2 gene encoded a valine and a tyrosine at amino acid positions 232 and 234, identical to the situation found in MEV type 1, but at position 300 there was a valine which is a determinant of MEV type 2. Immunization of mink with approximately 40,000 haemagglutinating units of recombinant MEV VP-2 induced a measurable antibody response as tested by haemagglutination inhibition. Furthermore, the immunized mink did not excrete virus and did not develop clinical disease upon challenge with a virulent isolate of MEV.
History
Journal
Journal of General VirologyVolume
75Pagination
149 - 155Publisher
Microbiology SocietyLocation
London, Eng.Publisher DOI
Link to full text
ISSN
0022-1317Language
engPublication classification
C1.1 Refereed article in a scholarly journalCopyright notice
1994, SGMUsage metrics
Categories
No categories selectedKeywords
AnimalsAntibodies, MonoclonalBaculoviridaeBase SequenceCapsidCells, CulturedCloning, MolecularFeline Panleukopenia VirusGenetic VectorsHemagglutination Inhibition TestsMinkMolecular Sequence DataVaccines, SyntheticScience & TechnologyLife Sciences & BiomedicineBiotechnology & Applied MicrobiologyVirologyCANINE PARVOVIRUSAUTONOMOUS PARVOVIRUSNUCLEOTIDE-SEQUENCEPROTEINSGENEMAP
Licence
Exports
RefWorks
BibTeX
Ref. manager
Endnote
DataCite
NLM
DC