Team:IOANNINA/Design
Design
Antibiotic deactivation module
Antibiotic Deactivation Module: This module is responsible for the enzymatic deactivation of tetracyclines and macrolides. It consists of the FAD-dependent monooxygenase TetX2 and the Erythromycin esterase EreB, expressed in a bicistronic form.
Antibiotic sensor
Antibiotic Sensor: The sensor we designed for the control of the state of “Dead” or “Alive” depends on the TetR and MphR transcriptional repressors. These genes are also expressed in a bicistronic mRNA molecule.
Kill switch
Kill Switch: Our kill switch which is a product of the partnership between our team and team IISER-Tirupati, consists of two parts, one part being a plasmid containing the protease mf-Lon in two copies, one under control of the macrolide dependent promoter pMph and one under the control of the tetracycline dependent pTet promoter. The second part of our kill switch is designed to be inserted in the bacterial chromosome and consists of the araC gene, bpDNase I modified with a tag for the recognition of the protein by mf-Lon, under the control of the arabinose inducible promoter.