FIG 5.

Determination of the presence of CheY4 in different mutant strains by immunoblotting. Total cell extracts obtained from cell cultures grown in Sistrom's minimal medium and harvested at an OD₆₀₀ of 0.6 were tested as described in Materials and Methods. Anti-CheY4 antibodies were used at 1:3,000. WT denotes the WS8N cell extract. Each lane is labeled according to the relevant mutation present in the strain. Other labels are the same as described in the legend for Fig. 4A. (B) Acrylamide gel electrophoresis of the RT-PCR products corresponding to the primer pair that amplifies cheY4 (lanes 1 to 4) and the primer pair that amplifies flgH (lanes 5 and 6). Lanes: 1, 3, and 5, total RNA from WS8N; 2, 4, and 6, total RNA from the fliA::kan mutant strain. Control reactions were carried out in the absence of reverse transcriptase (lanes 3 and 4). Lane MW corresponds to PhiX174 DNA digested with HaeIII and was used as a molecular size marker (fragment sizes, 1,353, 1,078, 872, 603, 310, 281, 271, 234, 194, and 118 bp). The expected sizes for the cheY4 and flgH products are 361 and 639 bp, respectively).