Subcellular localization of TraI.
A. Full-length TraI overexpressed in a wild-type strain from an inducible construct on the chromosome. TraI was detected by Western blot with TraI-specific polyclonal antiserum. Periplasmic (peri), membrane (memb), and cytoplasmic (cyto) fractions are indicated. A cross-reactive protein (∼35kDa) was found in the periplasmic fractions from strains with or without the T4SS. Antiserum to the gonococcal outer membrane protein PilQ was used to detect membrane material. Chloramphenicol acetyltransferase (CAT) was detected to identify cytoplasmic material.
B. Top Panels. Membrane fractions from wild-type (WT) and ΔGGI strains (left panel) and strains overexpressing (oe) traI or traIL6K/L12K from the native site (right panel). Bottom Panels. Cytoplasmic fractions from wild-type (WT) and ΔGGI strains (left panel) and strains overexpressing (oe) traI or traIL6K/L12K from the native site (right panel). TraI was detected by Western blot with TraI-specific polyclonal antiserum.