Journal article
Production of mink enteritis parvovirus empty capsids by expression in a baculovirus vector system: a recombinant vaccine for mink enteritis parvovirus in mink
The 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 baculo-viruses 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 40000 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.
Language: | English |
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Publisher: | Microbiology Society |
Year: | 1994 |
Pages: | 149-155 |
ISSN: | 14652099 and 00221317 |
Types: | Journal article |
DOI: | 10.1099/0022-1317-75-1-149 |
ORCIDs: | 0000-0002-5039-3178 |