Part:BBa_K2629001
Pseudomonas aeruginosa (Pyo) detector part - it activation allows the detection of 42bp of Pyo GyrA
This year, the goal of our team is to develop a fully automated system capable of : → identifying Pseudomonas aeruginosa → select the right phages for a possible phagotherapy
To do this, we will use a DNA probe: a plasmid with a one-stranded fraction which is complementary to the sequence we want to detect (target).
The target is a 42bp sequence found in Pyo GyrA gene. It encodes for the DNA gyrase subunit A (topoisomerase II). This chosen target is flanked with FspI and RsaI restriction sites, so its digestion is necessary to free the target fragment with correct length.
For our proof of concept, we worked on a pathogenic bacterial usually found in nosocomial infection cases due to its ability to develop antibiotic resistance : Pseudomonas Aeruginosa (Pyo), PAO1 strain. Here, is the description of the probe that characterize the lyse of Pyo which can detect a small fragment of its DNA. To do so, a plasmid containing the probe can be digestion with four specific restriction enzymes (cohesive and nicking enzymes) in order to create a single-stranded window. The feature is that the single-strand part will be specific of the target that we want to detect, this enable the recircularization of the plasmid.
For this part, the detector is inserted in psB1C3 containing BBa_J04450 allowing the emission of red fluorescence. Finally, the goal is to detect fluorescence in our automated system.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 53
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 10
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