Expression of MD infectious bursal disease viral proteins in baculovirus.
Genomic segment A of the variant infectious bursal disease virus (IBDV) strain MD was amplified by reverse transcriptase/polymerase chain reaction and further characterized by baculovirus expression. Three different baculovirus clones were constructed containing the genes encoding VP2, VP2/VP4, and the complete polyprotein cloned into the baculovirus transfer vector pVL1392. Baculovirus recombinants were identified by dot blot hybridization and were plaque purified three times. Baculovirus expression of the recombinants produced IBDV-specific proteins that were comparable in molecular size to native MD IBDV viral proteins VPX (48 kD), VP2 (45 kD), VP3 (32 kD), and VP4 (28 kD) as determined by sodium dodecyl sulfare-polyacrylamide gel electrophoresis and western immunoblot analysis. All three recombinants produced a 48-kD protein that possibly represents VPX, the precursor product of VP2. In addition to the 48-kD protein, the VP2/VP4 recombinant produced an IBDV-specific protein corresponding to the 28-kD VP4. The baculovirus-expressed polyprotein gene produced, in addition to the 48-kD protein, a 32-kD (VP3) IBDV-specific protein and a 29-kD protein that migrated slightly slower than MD VP4. The baculovirus-expressed proteins were used as antigens in an indirect enzyme-linked immunosorbent assay (ELISA). The ELISAs detected antibodies against the variant IBDV strains MD, GLS, and IN and the classic IBDV strains SAL and STC but did not detect antibodies against the variant Del-A and classic IBDV strain BVM or the serotype 2 IBDV strain OH.
Dybing
J K
JK
Department of Veterinary Preventive Medicine, Ohio Agricultural Research and Development Center, Ohio State University, Wooster 44691, USA.
Jackwood
D J
DJ
D000818:Animals; D016367:Baculoviridae; D002213:Capsid / Q000096:biosynthesis*; D036022:Capsid Proteins; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D016679:Genome, Viral; D007243:Infectious bursal disease virus / Q000235:genetics* / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection; D014771:Virion / Q000235:genetics / Q000378:metabolism
D036022:Capsid Proteins; D011994:Recombinant Proteins; C060339:VP4 protein, Rotavirus