Fig. 3.
The wt and NBS-deficient SecA2 proteins are expressed to equivalent levels in L. monocytogenes. (A) Summary of independently obtained recombinant L. monocytogenes clones that express chromosome-encoded wt (+) or NBS-mutated (−) SecA2-FLAG proteins. While wt SecA2-FLAG bacteria formed smooth (S) colonies, NBS-SecA2-FLAG bacteria formed rough (R) colonies on BHI plates. (B) Western blot analysis of SecA2 expression in three distinct wt or NBS-mutated bacterial clones encoding or not encoding (wt control) a C-terminal SecA2-FLAG protein. The cell pellet fraction of ∼3.3 × 108 log-phase bacteria was separated by SDS-PAGE, blotted, and revealed using a FLAG-specific MAb. The results shown are representative of two independent experiments that gave comparable results.