Extended Data Fig. 1: Characterization of functional metagenomic library delivery by hybrid T7 bacteriophage transducing particles into the target clinical strains.
a, Comparison of electroporation and transduction efficiencies. The figure shows that the maximum number of plasmids delivered to the hosts is at least two orders of magnitude higher by transduction than by electroporation in all 3 pathogenic host species (Centre and error bars represent mean and standard error (n= 3 biologically independent experiments)). Data available in Supplementary Table 2. b, PCR amplified metagenomic inserts from transduced cells. Following transduction and electroporation, the metagenomic DNA fragments were PCR amplified by using plasmid specific primers at both sides of the metagenomic DNA fragment and subsequently sequenced by capillary sequencing. This experiment differentiates monoclonal cells (single PCR product and DNA sequence) from those that were co-transduced by two or more plasmids (double bands on the gel and mixed signal in the capillary sequence). PCR was repeated in case of each host-library pair with similar results. c, During the generation of transducing bacteriophage particles, a large portion of phages remains replicative and kills the bacterial cells used for phage generation. Therefore, with the increasing phage concentration, transduction efficiency is not growing as would be expected, but declines. The figure shows the transduction efficiency of the T7 transducing phage particle harboring T7 phage tail (black line) on Shigella sonnei HNCMB 25021 at different dilutions (see Methods). Red dashed line shows the expected increase in transduction efficiency without any detectable killing effect of replicative phages. Data available in Supplementary Table 2.
