FIG 4.

RsiP degradation is dependent upon the site-2 protease RasP. B. thuringiensis wild type (EBT360) or ΔrasP (EBT366) containing a tetracycline-inducible copy of gfp-rsiP was subcultured 1:50 into LB supplemented with ATc (50 ng/ml). At mid-log phase, cultures were incubated with cefoxitin (5 μg/ml) for the time indicated at 37°C. The immunoblot was probed with anti-GFP antisera. EV is wild type with pAH9 (EBT169), and GFP is wild type with pAH13 (UM20). Streptavidin IR680LT was used to detect HD73_4231 (PycA homolog), which served as a loading control (62, 63). The color blot showing both anti-GFP and streptavidin on a single gel is shown in Fig. S6. Numbers at right are molecular masses in kilodaltons.