FIG. 6.

Expression and secretion of LCRS proteins in yscP Y. pestis. An immunoblot analysis of proteins expressed and secreted from Y. pestis KIM5-3001 (parent) and KIM5-3001.18 (ΔP2) or their Pla⁻ counterparts. KIM8-3002 (parent Pla⁻) and KIM5-3002.4 (ΔP2 Pla⁻), respectively, is shown. Bacteria were grown in TMH at 37°C without Ca²⁺. Proteins from soluble (s), membrane (m), and culture medium (e) fractions were separated by SDS-PAGE (12% polyacrylamide gel) and transferred to Immobilon P. Primary antibodies were used that were specific to YopE, YopD, YopH, YopM, LcrE, LcrQ, YscR, and V antigen (Vag), and the secondary antibody was conjugated to alkaline phosphatase. Arrows denote the positions of the respective proteins.