Expression and identification of the feline herpesvirus type 1 glycoprotein B (gp143/108).
The gene for feline herpesvirus type 1 (FHV-1) glycoprotein B (gB) has been cloned into an expression vector, pRVSVneo, containing the long terminal repeat of Rous sarcoma virus and polyadenylation signal of SV40. This expression vector containing FHV-1 gB gene, pRVSVgBneo, was transfected into Crandell feline kidney (CRFK) cells which are susceptible to FHV-1 infection. By indirect immunofluorescence analysis, the expressed gB was recognized with a panel of monoclonal antibodies (MAbs) against FHV-1 gp143/108. Immunoprecipitation analysis using a MAb 34H12 showed that molecular weights of the gB were 143 and 108 kDa under non-denaturing conditions that 108, 70, 64, and 58 kDa under denaturing conditions. The molecular weights were similar to those of the gB expressed in FHV-1-infected CRFK cells. In addition, when plasmid DNAs were injected into mice to obtain gB-monospecific serum, the pooled serum from mice inoculated with pRVSVgBneo, but not with pRVSVgDneo or pRVSVneo, recognized the FHV-1 gB polypeptides.
Maeda
K
K
Department of Veterinary Microbiology, Faculty of Agriculture, University of Tokyo, Japan.
Hirasawa
K
K
Kawaguchi
Y
Y
Ono
M
M
Mori
T
T
Gemma
T
T
Yokoyama
N
N
Doi
K
K
Mikami
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis; D002415:Cats; D002478:Cells, Cultured; D003001:Cloning, Molecular; D015870:Gene Expression; D005814:Genes, Viral; D005822:Genetic Vectors; D006564:Herpesviridae / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D014162:Transfection / Q000379:methods; D014611:Vaccination; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C078513:glycoprotein B, Felid herpesvirus 1