Stable implantation of orthogonal sensor circuits in Gram-negative bacteria for environmental release

Aitor de las Heras, Carlos A Carreño, Víctor de Lorenzo

Research output: Contribution to journalArticlepeer-review


A broad host range, orthogonal genetic platform has been developed to format sensor circuits in the chromosome of Gram-negative microorganisms destined for environmental release as bioindicators of toxic or perilous compounds (e.g. explosives) in soil. The genetic scheme includes the generation of a genomic landing pad for the sensor module with a Tn5-mini-transposon bearing an optimal attTn7 sequence and a choice of reporter systems with optical and enzymatic outputs. The array of functional elements thereby inserted in the chromosome match that of a cognate plasmid vector which delivers the transcription factors and the promoters to a frame that places the regulatory parts in front of the reporters. Site-specific recombination sites allow the deletion of antibiotic resistances and enables reporter output prior to deliberate release. The system thus allows the production and maintenance of cells in a pre-release state and its intentional conversion in deliverable strains that fulfil all safety, stability and performance criteria. The combination of such a genetic platform with a variant of the transcriptional regulator XylR of Pseudomonas putida that responds to 2,4-dinitrotoluene has been the basis for the production of strains that emit light upon exposure to residues of explosives in a soil microcosm.
Original languageEnglish
Pages (from-to)3305-16
Number of pages12
JournalEnvironmental Microbiology
Issue number12
Publication statusPublished - Dec 2008


  • Biosensing Techniques
  • DNA Transposable Elements
  • Dinitrobenzenes
  • Explosive Agents
  • Light
  • Plasmids
  • Pseudomonas putida


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