Plasmid construction and CMX expression for three different rBCG-CMX vaccines.

(A) PCR products corresponding to the CMX fusion gene, CMX (∼860 bp), from all three plasmid constructions and their respective empty controls: pLA71, pLA73 and pMIP12. M: molecular weight marker; NC: negative control reaction, using water; BCG: DNA from BCG-Moreau; rBCG transformed with pLA71, pLA73 and pMIP12, with (CMX) or without (empty) the fusion gene; PC: positive control reaction. (B) Analysis of CMX expression in rBCG-pLA71/CMX. Western blot of BCG transformants containing pLA71/CMX or empty vector using polyclonal antibody produced against rCMX. M: molecular mass marker; CMX: purified recombinant CMX; pLA71/CMX: rBCG with plasmid pLA71/CMX; pLA71: rBCG with plasmid pLA71.