Genome Integration and Excision by a New Streptomyces Bacteriophage, ϕJoe

• DOI: 10.1128/aem.02767-16
• Publication Date: 2016-12-22T02:11:48Z
• Authors: Paul C. M. Fogg; Joshua A. Haley; W. Marshall Stark; Margaret C. M. Smith
• Keywords: Integrases; Recombineering; Temperateness; Phagemid; Lysogenic cycle; Site-specific recombination

Bacteriophages are the source of many valuable tools for molecular biology and genetic manipulation. In Streptomyces, most DNA cloning vectors based on serine integrase site-specific recombination systems derived from phage. Because their efficiency simplicity, integrases also used diverse synthetic applications. Here, we present genome a new Streptomyces phage, ϕJoe, investigate conditions integration excision ϕJoe genome. belongs to largest phage cluster (R4-like) encodes integrase. The attB site venezuelae was efficiently by an integrating plasmid, pCMF92, constructed using int-attP locus. occupied in several genomes, including that S. coelicolor, mobile element varies gene content size between host species. Serine require phage-encoded directionality factor (RDF) activate reaction. RDF identified, its function confirmed vivo Both were active vitro assays. system is likely be important addition engineering toolbox.IMPORTANCEStreptomyces spp. prolific producers secondary metabolites, clinically useful antibiotics. Bacteriophage-derived engineering, as they enable heterologous into chromosome with ease high efficiency. Recently, researchers have been applying variety applications biology, rapid assembly novel combinations genes, biosensors, biocomputing. An requirement optimal experimental design predictability when integrases, however, need multiple enzymes different specificities sites. order provide broad platform identified validated newly isolated ϕJoe. specific recognition wide range actinobacteria, venezuelae, emerging model bacterium research.