17D yellow fever vaccine virus envelope protein expressed by recombinant baculovirus is antigenically indistinguishable from authentic viral protein.
We have constructed a recombinant baculovirus containing cloned DNA encoding the membrane and envelope (E) proteins of 17D yellow fever vaccine virus. Spodoptera frugiperda cells infected with this recombinant baculovirus produced a 66K protein which corresponded to the estimated size of the protein encoded by the cloned inserted DNA, and a 54K protein with the same molecular size as that of the authentic 17D yellow fever virus E protein. This recombinant 54K protein was labile, producing E protein-specific breakdown products (45K to 36K). Indirect immunofluorescence, using a panel of E protein-specific monoclonal antibodies, showed that the recombinant protein was presented both inside as well as on the surface of cells and was antigenically indistinguishable from the E protein of 17D yellow fever vaccine virus.
Shiu
S Y
SY
NERC Institute of Virology and Environmental Microbiology, Oxford, U.K.
Morikawa
S
S
Buckley
A
A
Higgs
S
S
Karunakarannair
V
V
Blachere
C
C
Gould
E A
EA
D000818:Animals; D000911:Antibodies, Monoclonal; D016367:Baculoviridae / Q000235:genetics; D002213:Capsid / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D000939:Epitopes / Q000032:analysis; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D015151:Immunoblotting; D008970:Molecular Weight; D009036:Moths; D011994:Recombinant Proteins / Q000276:immunology*; D015183:Restriction Mapping; D014614:Vaccines, Synthetic; D014758:Viral Core Proteins / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins / Q000235:genetics; D014765:Viral Vaccines; D015005:Yellow fever virus / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D004279:DNA, Viral; D000939:Epitopes; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins; D014765:Viral Vaccines