Table 2 Dinaciclib In silico HincII restriction pattern obtained for the 12,031 bp sequence spanning  wzi  to  gnd  in the Kp13  cps  gene cluster Start End Cut site Restriction fragment size between adjacent sites* (bp) 548 553 550   1,561 1,566 1,563 1,013 1,638 1,643 1,640 77 2,458 2,463 2,460 820 2,550 2,555 2,552 92 7,129 7,134 7,131 4,579 7,260 7,265 7,262 131 7,266 7,271 7,268 6 7,634 7,639 7,636 368 9,411 9,416 9,413 1,777 10,798 10,803 10,800 1,387 10,863 10,868 10,865 65 * Fragments used for this analysis are underlined. In vitro K-serotyping Kp13 Selleck Danusertib showed a weak positive reaction with both K9 and

K34 antisera that could not be resolved by modifying antiserum dilution or quellung reaction. This result is not surprising since cross-reactions

with the type-specific antisera is commonly observed among K. pneumoniae clinical isolates due to the activity of common genetic elements among distinct cps clusters [30]. In fact, the rmlBADC genes are also present in the cps cluster displayed by serotype K9 [15], and its CPS is composed of D-glucuronate, D-galactose and L-rhamnose residues [31]. Given the gene content of cps Kp13 and the presence of galE on the Kp13 genome, these residues could all be synthesized by this isolate, hence cross-reactions were not unexpected. From the comparison of cps Kp13 and cps check details VGH484 (K9, Figure 2) it is clear that they have common genes, but the Kp13 cps also has distinguishing features like its repertoire of GTs, the presence of uge-1 and a different cluster Chloroambucil organization (e.g. the positions of wzy and wzx). In the same line of evidence, the CPS of serotype K34 is composed of L-rhamnose, D-glucose and D-galacturonate residues [32], all of which also potentially present in the Kp13 CPS as discussed earlier, and D-galacturonate being produced by the epimerase activity from the uge-1 product. No cps sequences from K34 isolates were found on public databases. Nevertheless, our results indicate that Kp13 possess a unique

serotype since it showed a distinct RFLP pattern compared to those 102 patterns, including representatives of serotypes K9 and K34, previously described [29]. It has also been observed that cps-PCR genotyping seems to be a more sensitive and specific way for detecting novel serotypes [14], and our pyrosequencing-based approach together with the careful scrutinization of each CDS in the cluster and the in vitro results supports the finding that Kp13 synthesizes a novel CPS. Regulation of cps gene expression in Kp13 The transcriptional regulation of cps genes is thought to be under the control of three promoters, P1, P2 and P3, which are located upstream of galF, wzi and rmlB, respectively [13, 15]. As previously shown for other strains by Shu et al. [15], in the cps Kp13 cluster the transcripts driven by P1 and P2 should consist of galF/orf2 and wzi to gnd, respectively (Figure 4). Regulatory elements have been identified within the promoters P1 and P2 of the cps Kp13 cluster.