Part:BBa_K2629002:Design
Pseudomonas aeruginosa (Pyo) detector part - its activation allows the detection of 42bp of Pyo GyrA
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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 GyrA gene (2772bp) from PAO1 strain (GenBank : AAG06556.1) located in PA3168 locus. The target is located between nucleotide 229 and nucleotide 721. GyrA is found in all Pseudomonas aeruginosa strains and not linked in pathogenic behavior of Pseudomonas aeruginosa.