Fig. 4.

Construction of T1-spanin-loaded capsids and its bactericidal efficacy. (A) Schematic diagram of the construction of a φ80 phage capsid containing T1-spanin gene. T1-spanin exhibits toxicity toward synthetic bacterium 594; therefore, its expression was suppressed using a Tet repressor-expressing plasmid (pKLC83). Due to the presence of a packaging sequence, the T1-spanin plasmid was encapsulated within a phage capsid. (B) The synthesized phage capsid (A) was used to infect E. coli MC1061. Within bacterial cells lacking Tet repressor expression, T1-spanin was expressed. (C) E. coli MC1061 were infected with phage capsids containing either the T1-spanin-encoding plasmid or a plasmid lacking T1-spanin (control). The infected bacteria were plated on LB agar plates, and the number of colony-forming units (CFUs) was counted. Abbreviation: LB, Luria-Bertani medium. Assays were performed in triplicate. The bars show the means of the 3 spot test results, and the error bars show the standard deviations.