FIG. 2.

DUF582 proteins CT621, Cpn0852, CPn0726, and CT619 have TTS signals. The amino-terminal segments of the indicated proteins (24 to 30 amino acids, depending on the constructs) were fused to the Cya reporter protein and expressed in a Shigella flexneri ipaD (constitutive TTS) or mxiD (defective TTS) strain. Exponential-phase cultures expressing the reporter fusion protein were fractionated into supernatants (S) and pellets (P). Samples were resolved by SDS-PAGE, transferred to a PVDF membrane, and probed with anti-Cya (to detect chlamydial fusion proteins), anti-IpaD (Shigella secreted protein), or anti-CRP (Shigella nonsecreted protein) antibodies. All results shown are representative of at least two separate experiments.