Production of mink enteritis parvovirus empty capsids by expression in a baculovirus vector system: a recombinant vaccine for mink enteritis parvovirus in mink.

Abstract

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 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.

• Authors: J Christensen, S Alexandersen, B Bloch, B Aasted, A Uttenthal
• Journal: The Journal of general virology (J Gen Virol) Vol. 75 ( Pt 1) Pg. 149-55 (Jan 1994) ISSN: 0022-1317 [Print] England
• PMID: 8113722 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• Vaccines, Synthetic

Topics

• Animals
• Antibodies, Monoclonal
• Baculoviridae (genetics, metabolism)
• Base Sequence
• Capsid (biosynthesis, genetics, immunology)
• Cells, Cultured
• Cloning, Molecular
• Feline Panleukopenia Virus (genetics, immunology)
• Genetic Vectors
• Hemagglutination Inhibition Tests
• Mink (immunology, microbiology)
• Molecular Sequence Data
• Vaccines, Synthetic (biosynthesis, genetics, immunology)