Optimization of a whole-cell cadmium sensor with a toggle gene circuit.

This work demonstrates improvement of a whole-cell cadmium detection sensor through construction of a gene circuit. A cadmium (II) specific regulatory promoter, P(cadR,) from Psuedomonas putida 06909, is used in the assembly of a toggle circuit. The circuit contains the cadR promoter fused to lacIq and gfp, and a divergently transcribed tac promoter and cadR. The toggle sensor exhibits lower background fluorescence, and a 20-fold lower detection limit in comparison to a nontoggle gene circuit. The detection limit of the toggle sensor is 0.01 microM (1.12 ppb) cadmium chloride, and tunable with the addition of isopropyl-b-D-thiogalactopyranoside (IPTG). The toggle sensor is highly specific to cadmium (II), and no response is elicited from zinc, lead, manganese, nickel, copper, and mercury. 2009 American Institute of Chemical Engineers