High-level expression of canine parvovirus VP2 using Bombyx mori nucleopolyhedrovirus vector

For the potential use as recombinant vaccine, canine parvovirus (CPV) major capsid protein VP2 was expressed using Bombyx mori nucleopolyhedrovirus (BmNPV) vector. CPV VP2 gene was introduced into polyhedrin-based BmNPV transfer vector pBmKSK3, and recombinant virus BmK1-Parvo was prepared. When ant...

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Bibliographic Details
Published in:Archives of virology 2000-01, Vol.145 (1), p.171-177
Main Authors: CHOI, J. Y, WOO, S. D, LEE, H. K, HONG, H. K, JE, Y. H, PARK, J. H, SONG, J. Y, AN, S. H, KANG, S. K
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Biotechnology
Bombyx - virology
Bombyx mori
Canine parvovirus
Capsid - biosynthesis
Capsid - genetics
Capsid Proteins
Cells, Cultured
DNA, Recombinant - genetics
Dogs
Fluorescent Antibody Technique, Indirect
Fundamental and applied biological sciences. Psychology
Genetic Vectors
Health. Pharmaceutical industry
Hemagglutination Tests
Hemolymph - virology
Industrial applications and implications. Economical aspects
Nuclear polyhedrosis virus
Nucleopolyhedrovirus - genetics
Occlusion Body Matrix Proteins
Parvovirus
Parvovirus, Canine - genetics
Parvovirus, Canine - metabolism
Production of active biomolecules
Promoter Regions, Genetic
Recombinant Proteins - biosynthesis
Vaccins
Viral Proteins - genetics
Viral Structural Proteins
VP2 protein
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Summary:For the potential use as recombinant vaccine, canine parvovirus (CPV) major capsid protein VP2 was expressed using Bombyx mori nucleopolyhedrovirus (BmNPV) vector. CPV VP2 gene was introduced into polyhedrin-based BmNPV transfer vector pBmKSK3, and recombinant virus BmK1-Parvo was prepared. When anti-CPV.VP2 monoclonal antibody was employed in immunofluorescence staining, an intense signal was observed within BmK1-Parvo-infected Bm5 cells but not within uninfected cells or cells infected with a wild-type BmNPV-K1. In hemagglutination assay, the expression level of VP2 were 3.2 x 10(3) HA units/ml from infected Bm5 cells, 2.1x 10(5) HA units/larvae from infected larval fat body, and 1.6x 10(6) HA units/ml from infected larval hemolymph. These results suggested that BmNPV vector system using B. mori larva as host could be applied to efficient mass-production of recombinant vaccines.
ISSN:0304-8608
1432-8798
DOI:10.1007/s007050050014