Mapping of antigenic sites on the major inner capsid protein of avian rotavirus using an Escherichia coli expression system.
The cDNA encoding the VP6 gene of avian rotavirus PO-13 strain was inserted into the bacterial expression vector pET-3a. Upon isopropyl-1-thio-beta-D-galactoside induction, the E. coli BL21 (DE3) harboring the vector containing cDNA of the VP6 gene produced an approximately 45-kDa polypeptide, which reacted with rabbit serum against PO-13 strain in Western blotting. To study the antigenic sites on VP6, various deletion mutants were constructed, expressed in E. coli and the reactivity with antigenic site I- and II-specific MAbs analyzed by Western blotting. Site I, which is shared with all group A mammalian and avian rotaviruses except for chicken rotavirus, was found to be located at amino acid positions 45 to 65, and site II, which probably contributes to an authentic group A antigen common to both mammalian and avian rotaviruses, at amino acid positions 134 to 142.
Ito
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H
Department of Veterinary Public Health, Faculty of Agriculture, Gifu University, Japan.
Minamoto
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N
Goto
H
H
Luo
T R
TR
Sugiyama
M
M
Kinjo
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral; D000956:Antigens, Viral; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D003001:Cloning, Molecular; D018985:Epitopes, B-Lymphocyte / Q000032:analysis* / Q000276:immunology; D004926:Escherichia coli; D010957:Plasmids; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D012401:Rotavirus / Q000276:immunology*; D017384:Sequence Deletion
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D018985:Epitopes, B-Lymphocyte; D011994:Recombinant Proteins; C061332:VP6 protein, Rotavirus