FIG. 2.

Primer extension products from fla/che P_A and P_D-3. Total RNA from B. subtilis wild-type (LMB1), sigD null (LMB10), and flgM null (LMB213) strains that was isolated at the end of logarithmic growth (T₀) and 30 min later (T_0.5) was subjected to primer extension analysis. (A) Lanes 1 to 4, sequencing reactions of pWE1 (G, A, T, and C, respectively); lanes 5 to 8, primer extension products from total RNA, isolated by using primer OWE3, from B. subtilis LMB1 at T₀ (lane 5) and T_0.5 (lane 6) and from B. subtilis LMB10 at T₀ (lane 7) and T_0.5 (lane 8). (B) Lanes 1 to 4, sequencing reactions of pWE1 (G, A, T, and C, respectively); lanes 5 to 10, primer extension products from total RNA, isolated by using primer PD3, from B. subtilis LMB1 at T₀ (lane 5) and T_0.5 (lane 6), from B. subtilis LMB10 at T₀ (lane 7) and T_0.5 (lane 8), and from B. subtilis LMB213 at T₀ (lane 9) and T_0.5 (lane 10). The sequence given to the left of each panel corresponds to the nontemplate sequence, where +1 represents the nucleotide at which transcription initiates. Reactions were performed with 98 μg of LMB1 RNA isolated at T₀, 131 μg of LMB1 RNA at T_0.5, 153 μg of LMB10 RNA at T₀, 128 μg of LMB10 RNA at T_0.5, 84 μg of LMB213 RNA at T₀, and 92 μg of LMB213 RNA at T_0.5.