Figure 4.

Plasmid DNA co-purifies with MVs but is not transferred to E. faecalis or E. colicells. (A) DNA concentration was measured by Qubit in intact MV samples and in MVs lysed by boiling. Data shown from three independent experiments. Statistical analysis was performed by the unpaired t-test using GraphPad. ^****, P < 0.0001. (B) Agarose gel showing PCR product amplified with plasmid-specific primers on crude and purified MV fractions of WT and WT pGCP123. The expected plasmid PCR product is 240 bp. (C) Agarose gel showing PCR product amplified with plasmid-specific primers on intact and lysed MVs from WT pGCP123, subjected to DNAse treatment or treatment with inactivated DNAse prior to PCR. DNAse treated pGCP123 serves as a control. (D) The number of transformants following E. faecalis and E. coli incubation with MVs extracted from WT or WT pGCP123 determined by CFU enumeration on non-selective BHI or selective media with kanamycin. Statistical analysis was performed by the one-way ANOVA using one-way ANOVA test with Tukey's multiple comparison test. ^****, P < 0.0001.