Expression of the major inner capsid protein of the epizootic haemorrhagic disease virus in baculovirus and potential diagnostic use.
The RNA 7 encoding the major capsid protein (VP7) of epizootic haemorrhagic disease virus (EHDV), Australian serotype 2 (strain CS439), was cloned and the complete nucleotide sequence was determined. The coding region contained 1047 nucleotides (nt) capable of encoding a predicted 349 amino acid (aa) polypeptide with a calculated molecular size of 38.087 kDa. When the VP7 gene was expressed in bacterial or yeast expression systems, the expression product showed weak or no reactivity with polyclonal antibodies to EHDV. Therefore, the expression of the VP7 gene in baculovirus was pursued. The expressed EHDV VP7 was similar in antigenicity to that of the native virus as revealed by its reactivity in ELISA with monoclonal antibody (MAb) specific to EHDV. Preliminary ELISA results indicated that the recombinant protein binds to EHDV antibodies in serum and that these antibodies block the binding of EHDV-specific MAb. The availability of a reliable EHDV recombinant VP7 could enhance our existing assay for detection of EHDV-specific antibodies.
Nagesha
H S
HS
CSIRO Australian Animal Health Laboratory, Geelong, Victoria, Australia.
Gould
A R
AR
White
J R
JR
Lunt
R A
RA
Duch
C J
CJ
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D016367:Baculoviridae / Q000235:genetics; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D005822:Genetic Vectors; D018164:Hemorrhagic Disease Virus, Epizootic / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera / Q000166:cytology; D014709:Vero Cells
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C050779:VP7 protein, Rotavirus