FIG. 3.
Nucleotide sequence of the L. pneumophila iraAB locus. The deduced amino acid sequences of the various ORFs and the termination codons (*) are indicated. Arrows indicate the directions of translation of the ORFs. Putative promoter sequences are indicated by the −10 and −35 designations upstream of iraA, and the Shine-Dalgarno sequence is indicated as SD. The target sequence for the miniTn10 insertion in NU216 and NU216R is indicated in bold. The two underlined sequences represent a 250-bp IR. Restriction sites used for making the constructs described in this paper are indicated. The Kmr cassette insertion in NU244 is in the SacI site. The IraA sequences corresponding to the SAM-binding consensus and the IraB sequences corresponding to the PTR consensus are enclosed within parentheses. PCR was performed using primers 5′-AGCTAATCGTTTTGGTTCA and 5′-GCGCTACAGGAAAAGAATCAT to generate iraA-specific fragments and 5′CCGATGATGCCATTAAAGC and 5′TAACCAGCAGGGCGATAC to generate iraB-specific fragments for hybridization analyses. The primer-binding regions are indicated in lowercase type. For brevity, only the beginning of the upstream ORF is shown. Double-stranded sequence data were compiled with GeneRunner (Hastings Software, Inc., Hastings, N.Y.).