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Site-specific recombination of T2 phage using IP008 long tail fiber genes provides a targeted method for expanding host range while retaining lytic activity
Authors:Faezeh Mahichi  Aidan John Synnott  Keiko Yamamichi  Toshiya Osada  & Yasunori Tanji
Affiliation:Department of Life science, Tokyo Institute of Technology, Midori-ku, Yokohama, Japan;;Department of Bioengineering, Tokyo Institute of Technology, Midori-ku, Yokohama, Japan;and;Technical Department, Tokyo Institute of Technology, Midori-ku, Yokohama, Japan
Abstract:The application of bacteriophages (phages) in therapy urgently requires the production of wide-host-range recombinant phages that possess strong lytic activity. The wide-host-range IP008 phage was classified by transmission electron microscopy analysis as an A2 morphotype member of the Myoviridae family of the order Caudovirales . IP008 showed a high homology (99.4% similarity in the amino acid alignment of the major capsid protein Gp 23) with KEP10, another wide-host-range phage. The long tail fiber genes (genes 37 and 38 ) from the genome of T2 were replaced with those of the IP008 phage by homologous recombination. The host range of the recombinant phages was identical to that of IP008. Furthermore, the recombinant phage bacterial lytic activity was restored. Future analyses of host-range mutants of the closely related phages T2 and IP008 could lead to a more precise localization of the genetic factors responsible for receptor specificity.
Keywords:T-even bacteriophage  homologous recombination  tail fiber adhesion  host range
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