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Abstract
Bacteriophage vB_PaeP_PAO1_phiC725A (short name phiC725A) was isolated following mitomycin C induction of C7-25, a clinical Pseudomonas aeruginosa strain carrying phiC725A as a prophage. The phiC725A genome sequence shows similarity to F116, a P. aeruginosa podovirus capable of generalized transduction. Likewise, phiC725A is a podovirus with long tail fibers. PhiC725A was able to lysogenize two additional P. aeruginosa strains in which it was maintained both as a prophage and in an episomal state. Investigation by deep sequencing showed that bacterial DNA carried inside phage particles originated predominantly from a 700-800kb region, immediately flanking the attL prophage insertion site, whether the phages were induced from a lysogen or recovered after infection. This indicates that during productive replication, recombination of phage genomes with the bacterial chromosome at the att site occurs occasionally, allowing packaging of adjacent bacterial DNA.
Highlights
Some bacteriophages are capable of transduction, as shown by their capacity to transfer a DNA fragment from one bacterium to another, resulting in acquisition of new genetic information
As opposed to specialized transduction in which transduction is limited to host DNA adjacent to prophage insertion site [1], generalized transduction (GT) is defined as a process in which any host gene can be transferred to another bacterium
We demonstrate that the phage preferentially packages bacterial DNA localized on one side of its insertion site, when recovered by induction of the prophage, and after an infection cycle
Summary
Some bacteriophages are capable of transduction, as shown by their capacity to transfer a DNA fragment from one bacterium to another, resulting in acquisition of new genetic information. As opposed to specialized transduction in which transduction is limited to host DNA adjacent to prophage insertion site [1], generalized transduction (GT) is defined as a process in which any host gene can be transferred to another bacterium. Mu-like phages perform transduction following transposition at multiple sites in the bacterial chromosome, in the course of their replicative cycle [2, 3]. Other transducing phages perform rolling-circle replication and use packaging initiation (pac) sites for packaging their genome via a headful mechanism [4]. It has been suggested that cryptic pac site on the host chromosome may lead to packaging of bacterial DNA by such phages [5,6,7]. Phage F116, capable of GT, was isolated from a Pseudomonas
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