Production of infectious bacteriophage based on its genome is one of the necessary steps in the pipeline of editing phage genomes and creating synthetic bacteriophages. This process is called "rebooting" of the phage genome. In this chapter, we describe key steps required for successful genome "rebooting" using a native host or intermediate host. A detailed protocol is given for the "rebooting" of the genome of T7 bacteriophage specific to Escherichia coli and bacteriophage KP32_192 that infects Klebsiella pneumoniae.
Keywords: Gap repair cloning; Genome rebooting; Podovirus; Synthetic bacteriophage; Transformation-associated recombination cloning.
© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.