Determination of the ygjG transcriptional initiation site. Transcriptional initiation site was determined by primer extension. The reaction was carried out as described in Materials and Methods and cDNA transcript of RNA preparation obtained from cells grown under nitrogen limited conditions was analyzed on 6% PAGE (left track). The corresponding sequencing ladders are also labeled. The first nucleotide of ygjG mRNA (C) is bold and shown as "+1". The direction of transcription is shown by the arrow.