FIGURE 3.

Yeast‐based phage engineering. (1) The polymerase chain reaction purified fragments of phage genome along with fragments harboring gene inserts and reporter gene and a linear yeast artificial chromosome (YAC) vector with overhangs homologous to phage genome are (2) transformed into yeast cells. (3) The transformed phage DNA is assembled into the YAC vector through homologous recombination‐based Gap‐repair cloning. The recombinant YAC vector is amplified inside the yeast cells. (4) The modified YAC‐phage vector is extracted and purified. (5) The purified YAC‐phage vector upon transformation into host bacterial cells generates recombinant phage particles. (6) The resulting recombinant phage particles can be selected using a reporter gene (e.g., antibiotic resistance or fluorescence marker gene). The selected recombinant phage DNA is amplified into the bacterial host and released by cell lysis. The lysed recombinant phages can be used for various applications including targeted killing of MDR bacteria. Created using BioRender.com.