Mapping of the T and B cell epitopes of the Mycobacterium bovis protein, MPB70

Abstract

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 (γ‐IFN) production in a whole blood culture system. γ‐IFN production was measured using a sandwich enzyme immunoasssay specific ..