coli strains that cause cystitis The BLAST nucleotide algorithm (blastn) showed that pRS218 is 99% identical to plasmids pUTI89 [GenBank:CP000244], p1ESCUM [GenBank:CU928148] and pEC14_114 [GenBank:GQ398086] of E. coli causing acute cystitis, pUM146 [GenBank:CP002168] of a strain of E. coli associated with Crohn’s disease,
and pECSF1[GenBank:AP009379] of an E. coli strain belonging to the phylogenetic group B2 which was isolated from feces of a healthy adult (Figure 2) . Analysis of the repA1 sequence of FIIA replicon ACP-196 price of 24 IncFIB/IIA plasmids in pathogenic E. coli revealed three main lineages of virulence plasmids (Figure 3). All NMEC virulence plasmids were clustered into one lineage based on the repA1 sequence suggesting a common origin. Interestingly, pRS218 showed an identical origin with several virulence plasmids of E. coli causing cystitis (pUTI89 and pEC14_114), pECSF1 of the commensal MI-503 phylogenetic group B2 E. coli strain SE15 and pCE10A of NMEC strain CE10. Figure 2 Comparison of pRS218 sequence
to some virulence plasmids of other E. coli . Each code indicates a plasmid sequence. From top to bottom; pRS218, pUTI89 (a plasmid of the acute cystitis causing E. coli strain UTI89), pEC14_114 (a plasmid of
the uropathogenic E. coli strain EC14), pUM146 (a plasmid of the adherent invasive E. coli strain UM146), p1ESCUM (a plasmid of the acute cystitis causing E. coli strain UMN026) and pECSF1 (a plasmid of the commensal E. coli strain SE15). Each color box indicates clusters of ortholog genes present in plasmid sequences. White spaces in the blocks indicate the sequences that are not present in other plasmid sequences. Figure 3 Evolutionary relationship of IncFIB/IIA plasmids in pathogenic E. coli based on the repA1 sequence. The percentage of replicate trees in which the associated taxa diglyceride clustered together in the bootstrap test (500 replicates) is shown next to the branches. Genes of pRS218 are overly represented in NMEC strains compared to fecal E. coli Plasmid profiling revealed 27 of 53 (51%) of NMEC strains examined in the study harbored a plasmid similar in size to pRS218 (130-100 kb) (Table 2). Furthermore, PCR analysis revealed that a vast majority of pRS218-associated genes tested (n = 59) were overly represented (n = 52) among NMEC strains as compared to commensal E. coli (Table 3). Table 2 O serogroups of neonatal meningitis causing E.