Redesign of ultrasensitive and robust RecA gene circuit to sense DNA damage.

ABSTRACT: SOS box of the recA promoter, P_VRecA from Vibrio natriegens was characterized, cloned and expressed in a probiotic strain E. coli Nissle 1917. This promoter was then rationally engineered according to predicted interactions between LexA repressor and P_VRecA . The redesigned P_VRecA-AT promoter showed a sensitive and robust response to DNA damage induced by UV and genotoxic compounds. Rational design of P_VRecA coupled to an amplification gene circuit increased circuit output amplitude 4.3-fold in response to a DNA damaging compound mitomycin C. A TetR-based negative feedback loop was added to the P_VRecA-AT amplifier to achieve a robust SOS system, resistant to environmental fluctuations in parameters including pH, temperature, oxygen and

SUBMITTER: Chen JX 

PROVIDER: S-EPMC8601168 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature