Expression of a major bovine rotavirus neutralisation antigen (VP7c) in Escherichia coli.
Sequences from genomic RNA segment 8 of the United Kingdom tissue-culture (t.c.)-adapted bovine rotavirus encoding a major viral neutralisation antigen VP7c have been expressed in Escherichia coli. Expression under the regulated control of the bacteriophage lambda pR promoter was as a C-terminal extension to E. coli beta-galactosidase (beta Gal). Following temperature induction, high levels of the fusion protein were synthesised and accumulated in induced cells, making up 5%-15% of total bacterial cell protein after 2 h of induction. Immunisation of sero-negative rabbits and mice with gel-purified fusion-protein raised antibodies, which gave specific immunofluorescence with virus-infected cells and were able to immunoprecipitate proteins of the VP7 complex from such cells. Hyperimmune sera also gave a virus-type-specific reaction in a solid-phase enzyme-linked immunoabsorbant assay and neutralised virus infectivity in standard plaque-reduction assays.
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000941:Antigens / Q000235:genetics*; D000956:Antigens, Viral / Q000235:genetics*; D002417:Cattle; D011232:Chemical Precipitation; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D009500:Neutralization Tests; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012401:Rotavirus / Q000235:genetics*; D014614:Vaccines, Synthetic / Q000235:genetics*; D014765:Viral Vaccines / Q000235:genetics*
D000914:Antibodies, Viral; D000941:Antigens; D000956:Antigens, Viral; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014765:Viral Vaccines