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Mapping of the T and B cell epitopes of the Mycobacterium bovis protein, MPB70
Author(s) -
BillmanJacobe H.,
Radford A. J.,
Rothel J. S.,
Wood P. R.
Publication year - 1990
Publication title -
immunology and cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0818-9641
DOI - 10.1038/icb.1990.49
Subject(s) - epitope , biology , mycobacterium bovis , clone (java method) , microbiology and biotechnology , monoclonal antibody , antigen , recombinant dna , gene , epitope mapping , antibody , virology , immunology , genetics , mycobacterium tuberculosis , medicine , tuberculosis , pathology
Summary 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 for bovine γ‐IFN. B cell epitopes were mapped with a series of anti‐MPB70 monoclonal antibodies using an indirect enzyme immunoassay.