FIG. 2.

Sequence analysis of the cloned F. tularensis promoters. (A) Multiple alignment of all unique promoter sequences determined in this study. The promoters are arranged in ascending order. The TSP, −10, and −35 boxes were predicted using the BPROM tool and aligned using the T-coffee multiple-alignment tool. Nucleotides conserved among more than 50% of the sequences are indicated by black boxes. The predicted TSP nucleotide is boxed, while the 5 experimentally determined TSP nucleotides are circled. The consensus sequences of the −10, the −35, and the UP element are indicated at the bottom. The arrow on the bottom right indicates the TSP (nucleotide position +1). (B) Analysis of primer extension (PE) products specific for the indicated promoters. PE fragments were sized using GeneScan Analysis software (Applied Biosystems) (for detailed experimental conditions, see Materials and Methods). MapMaker 1000 internal lane standards are included in each electropherogram. The first 5′ nucleotide of the transcript is marked by an asterisk above the translation start diagnostic peak. The number above each panel refers to the length of the 5′ untranslated region of each promoter (i.e., the distance between the transcription starting point and the respective translation starting point). Note the absence of cDNA signal in the irrelevant-RNA negative-control panel.