Figure 2.

Construction of B. subtilis recombinant strain AZ703 expressing the chimera protein CotBΔ-BclA3_CTD. (A) Schematic representation of the strategy for the integration of the gene fusion cotBΔ::bcla3_CTD in the chromosomal DNA of B. subtilis PY79. The Western blot analysis of proteins extracted from spores of B. subtilis laboratory strain PY79 (lane 1 in B) and from the recombinant strain AZ703 (lane 2 in B) show bands of 66 and 46 kDa, which correspond to the endogenous CotB protein. The lane of recombinant strain AZ703 also displays a band of about 50 kDa which corresponds to the chimera protein CotBΔ-BclA3_CTD; 5 × 10⁸ of spores were resuspended in 100 µL of loading buffer and 20 µg of protein extract was loaded in SDS-PAGE gel. The immunoreaction was performed with anti-CotB antibodies and anti-rabbit secondary antibody conjugated with horseradish peroxidase.