Therefore, the objectives of the present study were to: (i) analy

Therefore, the objectives of the present study were to: (i) analyze the nucleotide sequence of pRS218 and its genetic and evolutionary relationship with virulence-associated plasmids

in other pathogenic E. coli, (ii) analyze the distribution of pRS218 genes among NMEC, and (iii) evaluate the contribution of pRS218 to NMEC pathogenesis GDC-0941 supplier by comparing the virulence of plasmid-cured and wild-type strains in vitro and in vivo. Results General properties of pRS218 Initial de novo assembly of short reads generated with Ion Torrent PGM technology identified 26 plasmid contigs ranging from 253 to 7,521 bp in length. These contigs were aligned to the reference plasmid sequence pUTI89 of uropathogenic E. coli strain UTI89 which was selected as the reference according to the sequence similarity of contigs (>90%). Complete sequence of pRS218 revealed that it is a circular plasmid of 114,231 bp in size with a G + C content of 51.02% (Figure 1). A total of one hundred and sixty open reading Mizoribine clinical trial frames (ORFs)

were annotated including IncFIB and FIIA replicons. Based on the blast analysis, nearly one third of the ORFs (n = 51) represents the genes involved in plasmid replication and conjugal transfer, along with 20 and 7 genes encoding mobile genetic elements

(MGEs) and products involved in DNA repair, respectively. Of the remaining ORFs, 59 encode unknown or hypothetical proteins, and 23 represent genes previously characterized in other bacteria. The plasmid does Decitabine not harbor any antibiotic resistance genes that may provide a selective advantage in the face of antibiotic therapy. Genetic load region of the pRS218 encodes several virulence- and fitness-associated genes which have been reported in other bacteria (Table 1). The annotated sequence of pR218 was deposited in GenBank at the NCBI [GenBank: CP007150]. Figure 1 Graphical circular map of pRS218. From outside to the center: ORFs in forward strand, ORFs in reverse strand, and GC skew. Plasmid genes are color coded as follows: Blue, conjugal transfer genes; Green, virulence or fitness-associated genes; Orange, plasmid replication genes; Red, IS elements; Black, plasmid stability genes; Light blue, hypothetical and putative genes. In the GC skew lime indicates the areas where the GC skew above Selleckchem Fosbretabulin average (51%) and purple indicates the areas below average.

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