FIG. 5.

Primer extension analysis of flaC transcription. The arrows indicate the surface-dependent flaC primer extension product. RNA was prepared from the wild-type strain BB22 or LM1017 after growth in HI broth (B), on HI plates (P), or in 2216 marine broth (2216B). Approximately 2 μg of total RNA was used in each primer extension reaction. Reactions 1 to 3 and 7 to 9 were primed with a flaC-specific oligonucleotide. Reactions 4 to 6 were primed with a flaD-specific oligonucleotide. Reactions 10 to 12 were primed with a flaB-specific oligonucleotide. Lanes: 1, BB22 (P); 2, BB22 (B); 3, LM1017 (P); 4, BB22 (P); 5, BB22 (B); 6, LM1017 (P); 7, BB22 (2216B); 8, BB22 (B); 9, BB22 (P); 10, BB22 (2216B); 11, BB22 (B); and 12, BB22 (P). Lanes g, a, t, and c correspond to the dideoxy nucleotide used in the sequencing reactions. Sequence and primer extension products were generated with flaC-, flaD-, or flaB-specific primers. The flaC-specific primer was 5′-CTGTTACAGCCATTTTGCTCTCC-3′.