title: Mapping of antigenic sites on the major inner capsid protein of avian rotavirus using an Escherichia coli expression system.
abstract: 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.
authors.affiliation: Department of Veterinary Public Health, Faculty of Agriculture, Gifu University, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Ito
authors.forename: N
authors.initials: N
authors.lastname: Minamoto
authors.forename: H
authors.initials: H
authors.lastname: Goto
authors.forename: T R
authors.initials: TR
authors.lastname: Luo
authors.forename: M
authors.initials: M
authors.lastname: Sugiyama
authors.forename: T
authors.initials: T
authors.lastname: Kinjo
mesh_terms: 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
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D018985:Epitopes, B-Lymphocyte; D011994:Recombinant Proteins; C061332:VP6 protein, Rotavirus