RECOMBINANT PHAGE AND METHODS
This disclosure provided methods of cloning a phage genome. Also provided are methods of making a recombinant phage genome. In some embodiments the phage genome is engineered to comprise a heterologous nucleic acid sequence, for example a sequence comprising an open reading frame. In some embodiments the phage genome is cloned in a yeast artificial chromosome. Recombinant phage genomes and recombinant phage are also provided. In some embodiments the methods are high throughput methods such as methods of making aa plurality of recombinant phage genomes or recombinant phage. Collections of recombinant phage genomes and recombinant phage are also provided.
1 . A method of making a cloned phage genome, comprising:
providing a vector;
inserting a starting phage genome into the vector to provide a recombinant vector; and
propagating the recombinant vector in a vector host cell that is not a phage host cell to thereby provide the cloned phage genome.
2 . A method of making a recombinant phage genome, comprising:
providing vector host cells comprising a recombinant vector comprising a cloned phage genome;
inserting a heterologous nucleic acid sequence into the starting phage genome to provide a recombinant phage genome; and
selecting vector host cells comprising the recombinant vector comprising the recombinant phage genome to thereby provide the cloned phage genome.
3 . A method of making a recombinant phage genome, comprising:
providing a yeast artificial chromosome comprising a cloned phage genome; and
inserting a heterologous nucleic acid sequence into the cloned phage genome to provide a recombinant phage genome.
4 . A method of making a phage, comprising:
providing a yeast artificial chromosome comprising a phage genome;
transforming the yeast artificial chromosome into competent phage host cells; and
isolating phage particles comprising the phage genome produced by the transformed phage host cells.