Immunogenicity and Protective Efficacy of a Novel Recombinant BCG Strain Overexpressing Antigens Ag85A and Ag85B
Figure 2
Recombinant Mycobacterium—E. coli shuttle plasmids (a) and expression of rBCG strains (b). The full-length copies of the M. tuberculosis fbpA(1.8 kb) or fbpB(1.5 kb) gene and its flanking regions including the promoter region were amplified from M. tuberculosis H37Rv chromosomal DNA, respectively. PCR products were digested by different enzymes and cloned into the Mycobacterium—E. coli shuttle vector pMV261. Ag85A and Ag85B proteins in the cell lysates and supernatants of rBCG::261, rBCG::85B, rBCG::85A, and rBCG::AB were detected by Western blotting analysis with anti-Ag85A or anti-Ag85B sera, respectively.
