ArticlePloS one2012
Directed evolution of the transcriptional regulator DntR: isolation of mutants with improved DNT-response.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
What it found
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Who cites it
9 citing papers in PubMed.
- Biological Switches: Past and Future Milestones of Transcription Factor-Based Biosensors.ACS synthetic biology · 2025Review
- Enhancing DNT Detection by a Bacterial Bioreporter: Directed Evolution of the Transcriptional Activator YhaJ.Frontiers in bioengineering and biotechnology · 2022Article
- Directed Evolution of Transcription Factor-Based Biosensors for Altered Effector Specificity.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Synthetic Biology Enables Programmable Cell-Based Biosensors.Chemphyschem : a European journal of chemical physics and physical chemistry · 2020Review
- Translating New Synthetic Biology Advances for Biosensing Into the Earth and Environmental Sciences.Frontiers in microbiology · 2020Review
- Tailor-made transcriptional biosensors for optimizing microbial cell factories.Journal of industrial microbiology & biotechnology · 2017Review
- Synthetic biology for the directed evolution of protein biocatalysts: navigating sequence space intelligently.Chemical Society reviews · 2015Review
- Genetically engineered microorganisms for the detection of explosives' residues.Frontiers in microbiology · 2015Review
- Transcription factor-based biosensors enlightened by the analyte.Frontiers in microbiology · 2015Review
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The transcriptional regulator DntR, which previously has been isolated from bacterial strains capable of degrading 2,4-dinitrotoluene (DNT), was engineered in order to improve the ability to detect DNT. A directed evolution strategy was employed, where sequence diversity first was created by random mutagenesis in three subsequent rounds, followed by recombination of previously selected mutants. A gfp gene was used as a reporter for transcriptional activity mediated by DntR and cells with higher GFP expression after addition of DNT were sorted out using fluorescence-activated cell sorting (FACS). A DntR mutant, which displayed 10 times higher induction levels than wild-type DntR in response to DNT was isolated. This mutant still maintained low levels of gfp expression in the absence of DNT. The detection limit was ∼10 µM, a 25-fold improvement compared to wild-type DntR. The functional role of some substitutions found in this clone is discussed in the framework of the structural changes observed when comparing the recently determined structures of DntR with and without bound inducer ligand.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.