TABLE 6.
Rescue of sporulation of CsgA
| Addition | Strain | Medium | Spores (%) |
|---|---|---|---|
| Crude C-factor from developing cellsa | DZ2 | MMC | 100 |
| DZ2 | Crude C-factor | 100 | |
| DK2657 (csgA) | MMC | 0.1 | |
| DK2657 (csgA) | Crude C-factor | 100 | |
| Ampicillin to CF saltsb | DK1622 | CF salts + ampicillin | 100 |
| DK2657 (csgA) | CF salts | ≤1 | |
| DK2657 (csgA) | CF salts + ampicillin | 33 | |
| Glycerol to CFc | DK1622 | CF | 100 |
| DK1622 | CF + glycerol | 100 | |
| DK2657 (csgA) | CF | ≤0.1 | |
| DK2657 (csgA) | CF + glycerol | 160 |
Rescue of sporulation of DK2657 (csgA) by crude C-factor. Crude C-factor was obtained from DZ2 developing in submerged culture as described in Materials and Methods. Freshly grown cells were then overlaid with the crude C-factor or MMC and allowed to develop for 1 week in submerged culture as described in Materials and Methods. Sonication-resistant spores were counted. The number of spores counted from the wild type developing in MMC was set to 100%.
Rescue of sporulation of DK2657 (csgA) by ampicillin. Strains were grown and prepared for development as described in Materials and Methods. Five-microliter aliquots of cells were spotted onto CF salts agar plates or CF salts agar plates containing ampicillin at 100 μg/ml. The plates were incubated at 28°C for 1 week. The spots of developing cells were harvested in 0.5 ml of water with a length of 7-mm-diameter glass tubing with a rubber bulb attached. The fruiting bodies were disrupted, and spores were released from the agar surface by sonication for 15 s at 30 W of power. The spores were counted in a hemacytometer and are reported as total spores per sample. The number of spores counted from the wild type developing on CF salts containing ampicillin was set to 100%.
Rescue of sporulation of DK2657 (csgA) by glycerol. Strains were grown and prepared for development as described in Materials and Methods. Five-microliter aliquots of cells were spotted onto CF agar plates or CF agar plates containing ampicillin at 100 μg/ml. The plates were incubated at 28°C for 1 week. The spots of developing cells were harvested in 0.5 ml of water with a length of 7-mm-diameter glass tubing with a rubber bulb attached. The fruiting bodies were disrupted, and the spores were released from the agar surface by sonication for 15 s at 30 W of power. The spores were counted in a hemacytometer and are reported as total spores per sample. The number of wild-type spores counted from CF plates was set to 100%.