Part:BBa_K2278001
Vibrio harveyi C8-CAI-1 (quorum sensing inducer) generator
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 136
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Introduction
1- Biological background
This DNA biobrick was designed in order to produce the C8-CAI-1of Vibrio harveyi in E. coli strain.The production of the Vibrio harveyi quorum sensing inducer (C8-CAI-1) is under the control of the cqsA gene coding for the CqsA synthase. This enzyme catalyzes the production of C8-CAI-1, an analog of CAI-1 quorum sensing inducer from V. cholerae. The CqsA catalyzes the following reaction :
2- Usage in iGEM projects
The BBa_K2278001 cames from the sensing module of the Croc’n cholera project (team INSA-UPS-France 2017) It was designed to produce C8-CAI-1 to simulate the presence of V. cholerae in a water sample and so, to allow the validation of our V. cholerae quorum sensing based detection system.
The part includes Vibrio harveyi’s cqsA synthase under the control of an IPTG inducible promoter. The C8-CAI-1 producing system is inducible in order to avoid toxicity problems and high metabolic activity during cells growth.
As the cqsA gene comes from V. harveyi which is a BSL1 organism, it can be used as substitute in experiment about the V. cholerae quorum sensing in BSL1 condition.
Characterization
3. Bioactivity of the C8-CAI-1 molecule
The molecule that we have produced has an activity in vivo because it activates the bioluminescence pathway of Vibrio harveyi JMH626. This vibrio strain used to detect C8-CAI-1 molecule because its deleted of other quorum sensing pathway and of the CqsA enzyme- as strong as the wild type strain or as our positive control (SN WT/JMH626). Still, a basal bioluminescence exist (Sn -/JMH626) but is really lower compared to SN with C8-CAI-1 (SN WT/JMH626).
None |