http://rdf.ncbi.nlm.nih.gov/pubchem/patent/WO-2022026446-A1

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publicationDate 2022-02-03-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber WO-2022026446-A1
titleOfInvention Methods of making unbiased phage libraries
abstract Described herein is a method of preparing an unbiased library of phage variants, comprising (a) preparing a population of "acceptor phage"; (b) removing an endogenous target gene and inserting gene variants into the acceptor phage genomes; (c) enriching the recombined phages; and (d) expressing the library for selection. The acceptor phage is a lytic phage comprising a synthetic genome wherein the target gene of interest is flanked by recoin binase sites. The acceptor phage infects a first host bacteria expressing a recombination plasmid facilitating recombination. The phages then infect a second host bacteria expressing a counterselection system that accumulates recombined phage variants and selecting against non-recombined phages. The accumulated phage variants infect a third host bacteria. The phage library may then be sequenced and characterized.
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