Synthesis in Escherichia coli of the major glycoprotein of human rotavirus: analysis of the antigenic regions.
Various regions of the gene encoding the major neutralization antigen, VP7, of human rotavirus have been expressed in Escherichia coli, as N-terminal fusions to beta-galactosidase under the control of the lac promoter. We have determined that the fusion products of two clones containing regions AB (aa 69-158) and ABC (aa 69-319) were antigenic, reacting with antibodies raised against whole virus. When guinea pigs were immunized with fusion protein purified by monoclonal antibody affinity columns, no neutralizing or virus-binding antibodies were detected, but antibodies binding to denatured VP7 were present.
Johnson
M A
MA
Department of Microbiology, University of Sydney, N.S.W., Australia.
Misra
R M
RM
Lardelli
M
M
Messina
M
M
Ephraums
C
C
Reeves
P R
PR
Bolcevic
Z
Z
Noel
J S
JS
Hum
C P
CP
Van Mai
H
H
D000956:Antigens, Viral; D015153:Blotting, Western; D002213:Capsid / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D036022:Capsid Proteins; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics*; D005814:Genes, Viral; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification; D015183:Restriction Mapping; D012401:Rotavirus / Q000235:genetics*; D015678:Viral Structural Proteins / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D000956:Antigens, Viral; D036022:Capsid Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins; C050779:VP7 protein, Rotavirus; D015678:Viral Structural Proteins; D001616:beta-Galactosidase