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Synchronization of Annd cultures. Synchronized growth of the HI-6 strain was obtained as in the work of Ghe and Ruby 23 with modifications. After two consecutive overnight subcultures 1: This resulted in seekinb synchronization: Synchronous growth lasted for about 11 h. Three samples were seeking at approximately 4 T1, growth initiation8 T2, midgrowth and 10 T3, septation, just before fragmentation h after the initiation ad growth, corresponding to optical density values at nm OD of 0. These samples are defined as stages of the growth phase GP. Synchronization of prey-dependent cultures. Synchronization of wt B. Under these conditions prey cells are attacked and penetrated synchronously as confirmed by phase microscopy.

This stage is followed by synchronous periplasmic growth of the invading cells. Cultures were sampled at 1, 2, and 3 h after mixing. A full cycle lasted for about 3. Bdelloplasts were harvested on a Percoll-sucrose cushion as described above. This protein fraction was run on two-dimensional gel electrophoresis 2DE gels see below. For the differential analysis, total cellular proteins from AP cells of wt and HI-6 strains and from synchronized GP stages of HI-6 were extracted with phenol according to the method of Barel et al. Each strip was then equilibrated twice in buffer according to the manufacturer's recommendations.

The gels were stained with Coomassie brilliant blue G 0. The strips were rehydrated with the protein samples for 1 h at zero voltage and then for 10 h at 50 V.

Equilibration was performed as described above. For comparative analysis between phases, one master gel was fot for each of the two pH ranges investigated. This was done by combining all the 2DE gel data for the relevant range. Within this set, only fo differing at least fivefold were selected for further analysis. Comparative analyses of the expressed proteome in the various cultures and time points were performed with at least three independent cultures. Tryptic peptides were resolved by reverse-phase chromatography on 0.

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