To build our engineered gut probiotics system, we constructed four plasmid systems that collectively can be stably expressed in ECNs. They are pMUT1-G_Exothermic suicide, pMUT2mini_quorum sensing system, pFB3_Hydrogel expression system and pFB1_Synthetic feedback system to achieve the functionality we want to achieve. These plasmids have high stability and are not easy to lose in a non-resistant environment, which can ensure the stable functioning of intestinal probiotics for a long time. At the same time, we also removed the oriT region and Nika fragment in the plasmid vector pMUT1-G while ensuring the normal function of the plasmid, so that the plasmid could not be transferred horizontally.
