Biosynthetic Trigger System Based On LuxI/R QS Signals

Triggers are widely applied in the field of fermentation industry, environmental monitoring and food industry. Based on gene circuit and gene engineering, synthesis biology provides a conductive foundation for the design and construction a biotrigger system in vivo. Combining with the conception of synthetic biology and methods of molecular biology, this study constructed in silico and in vivo an E.Coli TOP10 biotrigger based on LuxI/R QS Signals pathway. The results are as follows:1. An artificial biotrigger system based on LuxI/R is firstly designed and simulated by mathematical model. The results show that the strength of promoters, generation Rate and degradation of related proteins will effect on the sensitivity and stability of the system. In different system state, the system will response to the input signal and get to the only final status.2. The biotrigger related functional gene component, including luxI generator and AHL receptor are constructed and ligated by molecular biological method. LuxI generator are tested to produce 3OC6HSL signal molecular by LC-MS-MS; further fluorescence research indicates AHL receptor could respond to AHL signal, and produce green fluorescence protein(GFP).3. Further study displays that the biotrigger could only respond and express GFP when input signal, IPTG concentration is changing. And the response range of IPTG concentration is from 0.01mM to 0.5mM, which indicates the sensitivity of this system is good. And this system is able to response to IPTG signal in different cell growth phase(From beginning to OD0.6).