Part:BBa_K2629000:Design

The goal of our detector is to reveal the presence of Pseudomonas aeruginosa in a sample thanks to the specific lysis by a bacteriophage. The design of this detector has been inspired by Cork Ireland 2015 team : they created different based on the perfect complementarity of the double strand DNA.
Thus, we - Grenoble team 2018 - decided to create a similar detector for Pseudomonas aeruginosa pathogen.

Design Notes

The probe is designed in order to detect a fragment less than 100 bp. The aim is to create a single strand window that enable the perfect hybridization of our choosen target. It is made by: → Two restrictions enzymes producing cohesives end, SphI and NgoMIV, which goal is to remove the little sequence in between on the bottom strand and thus create a perfect complementarity with the target. But at the end we trust that using PCR linearization could reduce the background of uncut plasmid. → Twi nicking enzymes, Nt.BspQ1 and Nb.BssS1, enzymes that cut one strand of the double DNA strand.

Source

The aim is to detect a small fragment of Pseudomonas aeruginosa DNA. The target was found in ProC gene (822bp) from PAO1 strain (GenBank : AAG03782.1) located in PA0393 locus. The target is located between nucleotide 766 and nucleotide 802. ProC is an housekeeping gene found in all Pseudomonas aeruginosa strains and not linked in pathogenic behavior of Pseudomonas aeruginosa.