Something Interesting

We decided to use the vector pUCP18 as a starting point to build our plasmid. It contains the origin of replication pRO1600 oriV BBa_K4083013 which originates from Pseudomonas aeruginosa POA and enables plasmid amplification in various Gram-negative bacteria [1]. Additionally, a high copy number origin of replication, ColE1 BBa_K4278506, is included in the backbone. This allows efficient plasmid replication and maintenance in E. coli [2], facilitating cloning procedures and plasmid manipulation.

The promoter is the most important building block to restrict antimicrobial peptide production to the targeted bacteria. CAPTURE uses a mutated version of the P. aeruginosa specific pel promoter.

In addition to the constitutive pel promoter, we also needed an inducible promoter for testing in the lab. We settled on the natural, Pseudomonas specific and xylose-inducible P_xut promoter, which is already characterized in Pseudomonas aeruginosa as well as in Pseudomonas fluorescens [7].

The heart of the plasmid is the gene encoding an antimicrobial peptide which is produced by the target pathogen Pseudomonas aeruginosa, eventually leading to the death of the bacterial cell.