BioDesign Research / 2020 / Article / Fig 1

Research Article

Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All

Figure 1

PICI assembly and rebooting workflow. For the assembly of a YAC-PICI, a PCR fragment containing the yeast replicon and selection marker from pAUR112 is required. Additionally, PCR products containing the PICI regions to assemble in the YAC vector by gap-repair cloning are used. All PCR fragments present, at their ends, partially overlapping regions. The final product will mirror the PICI structure in the chromosome. YAC-PICI DNA is extracted from the yeast and directly transformed into competent cells of the host bacterium. Then, the PICI excises from the YAC and integrates into its corresponding attachment site located on the chromosome. The whole process can be followed through the selective marker present in the island. For rebooting the island, the induction of its cognate helper phage is needed.