Expression of the p80 region of bovine viral diarrhea virus and identification of specific antibodies to this recombinant protein in bovine sera.
A 917-base pair segment of the bovine viral diarrhea virus (BVDV) genome encoding part of the p80 region was cloned into plasmid Gex-2T expression vector for expression as a fusion protein with glutathione-S-transferase (GST). When the p80 and GST sequences were in the same reading frames, the resulting GST-p80 fusion protein had a molecular mass of 58 kilodaltons (kDa) in SDS-PAGE. Extracts of control E. coli carrying only the vector plasmid (Gex-2T) did not contain this new 58-kDa protein band. Mouse monoclonal antibody specific to BVDV-p80 recognized this recombinant protein. Seventy cattle sera that had an SN titer (to TGAC isolate of cytopathic BVDV) greater than 1:8 reacted with this recombinant protein in Western blots. Of 28 cattle sera that had SN titers less than or equal to 1:8, only one serum tested positive on Western blots.
Kwang
J
J
USDA, ARS, U.S. Meat Animal Research Center, Clay Center, NE 68933.
Bolin
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S
Littledike
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ET
Dubovi
E J
EJ
Donis
R O
RO
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D015153:Blotting, Western; D002417:Cattle; D003001:Cloning, Molecular; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D005982:Glutathione Transferase / Q000235:genetics / Q000302:isolation & purification; D008970:Molecular Weight; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D005982:Glutathione Transferase