Bordetella bronchiseptica bacteriophage deploys a combinatorial library approach to the challenge of maintaining infectivity to a highly variable host bacteria.  The virus maintains a vast library of potential tail fiber proteins to bind to the ever-changing surface of its host. The phage-encoded reverse transcriptase, using a non-coding region as a template, generates trillions of mutations across a 44-residue variable region required for phage attachment to its bacterial host. We have demonstrated the viability of this system by successfully identifying a binder to a specific target with the tail fiber protein of the virus. Furthermore, we describe the insertion of heterologous sequences into the template region, and phage-based mutagenesis for library generation.  Expanding the naturally occurring diversity generating mechanism allows straight-forward access to libraries that could greatly exceed the diversities of conventional molecular display techniques.

Self-made phage libraries optimized for target binding

243rd National Meeting (2012)

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