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. 2020 Feb 18;5(1):e00684-19. doi: 10.1128/mSystems.00684-19

FIG 8.

FIG 8

Genomic organization and PCR validation of SpST68A. (A) Chromosomal region of an ST68 genome (NCBI GenBank accession CP015626) with integrated SpST68A prophage shown in red. As indicated, the SpST68A is inserted between a tRNA locus (locus tag A6M57_13930) and a hypothetical gene (locus tag A6M57_8065). A non-ST68 genome (NCBI GenBank CP016073) without the SpST68A prophage is shown here for comparison. The P1F, P3F, and P2R primers used for PCR screening of SpST68A are shown as small black arrows. (B) Schematic diagram showing the SpST68A insertion site and PCR amplification strategy in a greater detail. (C) A representative DNA agarose gel showing PCR screening of SpST68A prophage using P1F/P2R and P3F/P2R primer pairs. Both set of PCRs were performed on all 50 S. pseudintermedius isolates sequenced in this study. As shown, P1F/P2R PCR could not amplify the 46,523-bp region (SpST68A) expected in ST150 and ST1049 but amplified a 980-bp region in isolates lacking SpST68A. P3F/P2R PCR amplified the 1,491-bp region in ST150 and ST1049 but not in other isolates.