Mapping of the T and B cell epitopes of the Mycobacterium bovis protein, MPB70.
A clone coding for the entire gene for the Mycobacterium bovis protein antigen MPB70 was used to produce a series of overlapping subclones by making a series of deletions from the 3' end of the gene. The subclones expressed incomplete MPB70 proteins as fusions with glutathione-S-transferase. The insert DNA was sequenced to determine the extent of the deletion and the proteins expressed by the clones were examined for the presence of T cell and B cell epitopes. T cell epitopes were mapped by measuring the ability of recombinant antigens to stimulate gamma interferon (gamma-IFN) production in a whole blood culture system. gamma-IFN production was measured using a sandwich enzyme immunoassay specific for bovine gamma-IFN. B cell epitopes were mapped with a series of anti-MPB70 monoclonal antibodies using an indirect enzyme immunoassay.
Billman-Jacobe
H
H
CSIRO Division of Animal Health, Animal Health Laboratory, Parkville, Victoria, Australia.
Radford
A J
AJ
Rothel
J S
JS
Wood
P R
PR
D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D002417:Cattle; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D005982:Glutathione Transferase; D007371:Interferon-gamma / Q000032:analysis; D008969:Molecular Sequence Data; D009154:Mutation; D009163:Mycobacterium bovis / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins; C060216:MPB70 protein, Mycobacterium bovis; D007371:Interferon-gamma; D005982:Glutathione Transferase