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 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.
Choi
J Y
JY
Division of Applied Biology and Chemistry, Seoul National University, Suwon, Korea.
Woo
S D
SD
Lee
H K
HK
Hong
H K
HK
Je
Y H
YH
Park
J H
JH
Song
J Y
JY
An
S H
SH
Kang
S K
SK
D000818:Animals; D012831:Bombyx / Q000821:virology*; D002213:Capsid / Q000096:biosynthesis* / Q000235:genetics*; D036022:Capsid Proteins; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000235:genetics; D004285:Dogs; D019084:Fluorescent Antibody Technique, Indirect; D005822:Genetic Vectors; D006386:Hemagglutination Tests; D006458:Hemolymph / Q000821:virology; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D000077183:Occlusion Body Matrix Proteins; D017993:Parvovirus, Canine / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins
D036022:Capsid Proteins; D004274:DNA, Recombinant; D000077183:Occlusion Body Matrix Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus