FIG. 4.

Determination of the transcriptional start site by primer extension analysis. Total RNAs (40, 10, 10, and 10 μg) from B. subtilis 168 cells cultured in DSM at t_0.5, DSM with 2.5 mM maltose [DSM (Mal)] at t₋₁ and t_0.5, and mSMM at t_0.5, respectively, were used as RNA samples. Signals were detected with ³²P-labeled primer V1-PEX. Dideoxy DNA sequencing reaction mixtures with the same primer were electrophoresed in parallel (lanes G, A, T, and C). The nucleotide sequence of the transcribed strand is given beside the sequence ladder and the arrow indicates the nucleotide at the transcriptional start site. A map of the glv operon and the nucleotide sequence of the upstream region of glv are shown below the primer extension analysis results.