FIG 2.
Synthesis and localization of the diadenylate cyclase CdaA. (A) Expression of cdaA in cdaR and cdaA mutant strains of L. monocytogenes was analyzed by Western blotting. The protein extracts of strains EGD-e (wild type), LMJR45 (ΔcdaR), and LMJR46 (IcdaA) grown with or without IPTG were separated by SDS-PAGE, and the CdaA levels were visualized using a polyclonal antiserum specific for L. monocytogenes CdaA. (B) Effect of cdaR inactivation on CdaA membrane localization. Strains EGD-e (wild type) and LMJR45 (ΔcdaR) were grown in BHI medium at 37°C, and the bacteria were harvested at an OD600 of 1.0. Cytosolic (C, 15 μg) and membrane (M) fractions of each strain were separated by SDS-PAGE (12%) and analyzed by Western blotting using CdaA antiserum. The protein labeled by an asterisk might be CdaA that is processed due to overproduction of the protein.