Part:BBa_K2278002

Introduction

This DNA biobrick was designed in order to produce CAI-1 of Vibrio cholerae in E. coli strain.

1- Biological background

The production of the Vibrio cholerae quorum sensing inducer (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 Vibrio cholerae. The CqsA catalyzes the following reaction :

Figure 1: CAI-1 simplified production mechanism. CqsA synthase catalyses the reaction between (S)-adenosylmethionine (SAM) and deca-coenzyme A. Ea-CAI-1 is then converted to CAI-1 (Wei et al. 2011). (figure is adapted from Wei et al. 2011)

2- Usage in iGEM projects

The BBa_K2278002 cames from the sensing module of the Croc’n cholera project (team INSA-UPS-France 2017) It was designed to produce CAI-1 by an E.coli to simulate the presence of Vibrio cholerae in a water sample and so, to allow the validation of our Vibrio cholerae quorum sensing based detection system.

The part includes Vibrio cholerae cqsA synthase under the control of an IPTG inducible promoter. The 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 cholerae which is a BSL2 organism, it has to be manipulated under safe condition even if this gene is not directly involved in pathogenicity. To avoid safety issues we recommand to use BBa_K2278001 which produce a quorum sensing analog molecule : C8-CAI-1