Fig 1.
Schematic map of the native plasmid pHS87a and pHS87b found in the P. aeruginosa clinical isolate HS87.
Mobile elements are indicated by the lines above or below the sequence. Antibiotic resistance gene is black; genes with predicted functions are grey; function unknown genes are white. The numbers are orf numbers of the function unknown genes. The black triangles facing each other at the termini of a line indicate inverted repeats (IRs) of the element. The single triangle appearing on the line of ΔIS1071 means it is a truncated element. Direct repeats flanking the elements are shown by corresponding letters. Syntenic regions are linked by grey parallelograms or squares with identities shown. The schematic is drawn to scale.
Fig 2.
Site-specific integration of pHS87b in to the tRNAThr gene site.
(A) Integration schematic. Black triangles indicate homologous attachment regions and direct repeats. Genes are indicated by thick arrows. Primers are shown as thin arrows. (B) tRIP-PCR at the assay tRNAThr gene site showed a faint band. Amplification of 16S rRNA gene and attachment site of pHS87b was also performed. (C) Amplification of junctions of the integrated pHS87b. (D) Sequencing confirmed the integration of pHS87b into the tRNAThr gene site. The tRNAThr gene sequence is underlined. The block highlights the homologous region. (E) Alignment of the integrated form of pHS87b with the syntenic chromosomal regions of P. aeruginosa NCGM 1984 and NCGM 1900. The schematic is not drawn to scale.