Part:BBa_K2278001:Design
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]
This DNA biobrick was designed in order to produce C8-CAI-1 of 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.
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 Vibrio harveyi in a water sample and so, to allow the validation of our Vibrio harveyi quorum sensing based detection system, as first step toward Vibrio cholorae detection.
The part includes Vibrio harveyi 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 Vibrio harveyi which is a BSL1 organism, we used it as a substitute to the pathogenic Vibrio cholerae
Sequencing revealed that the VhCqsA construction slightly differs from the initial design, with a loss of the 9 last amino acid of the protein (position 382 to 391). This does not prevent its functionality, as demonstrated in the main page section.Source
This enzyme came from Vibrio harveyi DNA genomic sequence.
References
Henke, J. and Bassler, B. (2004). Three Parallel Quorum-Sensing Systems Regulate Gene Expression in Vibrio harveyi. Journal of Bacteriology,186(20), pp.6902-6914. Sequence is available at: https://www.ebi.ac.uk/ena/data/view/AAT86008&display=text