FIG. 8.
Analysis of Yop synthesis and secretion from Y. pseudotuberculosis strains grown either with (+) or without (−) Ca2+. Yops in the total protein fraction (a mix of proteins secreted to the culture medium and contained within intact bacteria) (upper panel) or secreted to the extracellular medium (cleared culture supernatants) (lower panel) were separated by SDS-PAGE and identified by immunoblot analysis using a polyclonal rabbit antiserum recognizing secreted Yops. Where indicated, IPTG was added at a final concentration of 0.4 mM upon temperature shift. Lanes: a and b, wild-type YPIII/pIB102; c and d, yscX null mutant YPIII/pIB880; e and f, complemented YPIII/pIB880, pJEB291 (YscX+); g and h, complemented YPIII/pIB880, pJEB295 (Pcr3+); i and j, yscY null mutant YPIII/pIB890; k and l, complemented YPIII/pIB890, pJEB292 (YscY+); m and n, complemented YPIII/pIB890, pJEB296 (Pcr4+). Molecular masses shown in parentheses are deduced from primary sequences.