The G+C value for each orf in MGH 78578 is shown below each orf. The red bar indicates the corresponding location replaced by an apramycin resistant gene in the promoter knocked-out strain, NK8-Δcit, derived from the NK8 clinical strain. Corresponding citrate fermentation loci from S. enterica serovar Thiazovivin purchase Typhimurium LT2 and E. coli K12 are shown (b and c)

with colours indicating homologous genes. Alternative gene names in parentheses on top of some orfs for better comparison were based on homology search. The locations of these regions in the genomes are marked below. In the LT2 genome, two clusters of citrate fermentation genes were found. The corresponding flanking genes for locus I, dcuC and rna, and locus II, rihC and dapB, are shown in black. Another gene cluster containing the citWX and the divergent

citYZ genes are conserved among K. pneumoniae genomes (Figure 1a). In NTUH-K2044, ARRY-438162 molecular weight the citWX-citYZ gene cluster is located at 15,693-bp downstream of the dapB. The existence of this additional gene cluster, especially the citX, is important for the function of citrate lyase in K. pneumoniae. Unlike the counterpart identified in Salmonella enterica (Figure 1b), the 13-kb region in K. pneumoniae does not contain citX for the biosynthesis of the prosthetic group of citrate lyase [7]. In MGH 78578, the deduced amino acid sequences of citY and citZ are 43% and 41% identical to CitA and CitB, respectively. Nearly 4EGI-1 manufacturer half of the K. pneumoniae clinical isolates carry the 13-kb genomic island The presence/absence of the 13-kb region was investigated in additional K. pneumoniae clinical isolates (NK3, NK5, NK6, NK8, NK9, NK25, NK29, NK245, CG43, CMKa01 through CMKa08, Celecoxib CMKa10). These isolates were collected from patients with pneumonia (3), bacteremia (4), liver abscess (7), UTI (2), meningitis (1), and endophthalmitis (1). We conducted comparative genomic hybridization (CGH) analysis on the test strains with custom-made DNA

microarray (NimbleGen), in which a total of 389,266 probes were designed based on the CDSs of five sequenced K. pneumoniae genomes [12]. For the current report, we have analyzed the results of the predicted coding sequences spanning the 13-kb region of MGH 78578. As shown in Figure 2, each of the 19 strains (including MGH 78578 as a control) was compared against the NTUH-K2044 reference genome. The dots represent the DNA copy number log ratios between the reference and tested genomes for the 687 probes corresponding to the sequences spanning the 13-kb region. Since the NTUH-K2044 genome does not carry the cit genes, these results indicate that the 9 strains with dots plotted at the baseline in this region (NK5, NK6, NK9, CG43, CMKa01, CMKa02, CMKa04, CMKa08, and CMKa10) do not carry these genes in their genomes. The other ten strains shown in below, including MGH 78578, gave higher signals for the cit genes than that from the reference (Figure 2).