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. 2023 Mar 22;11(2):e05179-22. doi: 10.1128/spectrum.05179-22

FIG 5.

FIG 5

Biofilm-derived P. aeruginosa PAO9503 OMVs are more efficient at mediating horizontal gene transfer of plasmid DNA encoding antibiotic resistance than planktonic-derived P. aeruginosa PAO9503 OMVs. (A) The number of transformants resulting from transformation with either 2 × 109 planktonic-derived PAO9503 pBBR1MCS-5 OMVs (pOMVs; closed circles), control non-treated PAO9503 pBBR1MCS-5 biofilm-derived OMVs (bOMVs; filled squares), DNase-treated PAO9503 pBBR1MCS-5 biofilm-derived OMVs (bOMVs + DNase; open squares), or 500 ng of plasmid DNA alone (filled triangles) are shown. Data shown are means ± SEM of n ≥ 3 biological replicates. ns, not significant; *, P < 0.05 (one-way ANOVA with Tukey’s multiple comparisons). (B) PAO9505 transformants resulting from transformation using either non-treated P. aeruginosa PAO9503 biofilm-derived OMVs or PAO9503 planktonic-derived OMVs, both harboring pBBR1MCS-5, were confirmed by performing colony PCR using primers specific for the pBBR1MCS-5 gentamicin cassette. A positive plasmid control transformant colony (pos. control col.) and 1 ng pBBR1MCS-5 DNA (PCR + control) were used as positive controls. A non-transformed PAO9505 colony was used as a negative control for the PCR (PCR - control). Arrow indicates the gentamicin cassette amplicon (Gmr; 500 bp). Image is representative of n = 4 biological replicates.