ArticleAMB Express2020
Directed evolution of a transcription factor PbrR to improve lead selectivity and reduce zinc interference through dual selection.
Article in AMB Express, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Active learning-guided optimization of cell-free biosensors for lead testing in drinking water.Nature communications · 2025Article
- Biological Switches: Past and Future Milestones of Transcription Factor-Based Biosensors.ACS synthetic biology · 2025Review
- Systematic evaluation of pigment-based whole-cell lead biosensors: challenges in genetic circuit engineering and critical considerations for background noise control.Frontiers in bioengineering and biotechnology · 2025Article
- Bacterial Metallostasis: Metal Sensing, Metalloproteome Remodeling, and Metal Trafficking.Chemical reviews · 2024Review
- Engineering a cell-free biosensor signal amplification circuit with polymerase strand recycling.bioRxiv : the preprint server for biology · 2024Article
- Directed Evolution of 4-Hydroxyphenylpyruvate Biosensors Based on a Dual Selection System.International journal of molecular sciences · 2024Article
- Steering and controlling evolution - from bioengineering to fighting pathogens.Nature reviews. Genetics · 2023Review
- Advances in Synthetic-Biology-Based Whole-Cell Biosensors: Principles, Genetic Modules, and Applications in Food Safety.International journal of molecular sciences · 2023Review
- Pb(II)-inducible proviolacein biosynthesis enables a dual-color biosensor toward environmental lead.Frontiers in microbiology · 2023Article
- Synthetic bacteria for the detection and bioremediation of heavy metals.Frontiers in bioengineering and biotechnology · 2023Review
- Metabolic engineering of the carotenoid biosynthetic pathway toward a specific and sensitive inorganic mercury biosensor.RSC advances · 2022Article
- The protein scaffold calibrates metal specificity and activation in MerR sensors.Microbial biotechnology · 2022Article
- Directed evolution of the PobR allosteric transcription factor to generate a biosensor for 4-hydroxymandelic acid.World journal of microbiology & biotechnology · 2022Article
- Engineering of Synthetic Transcriptional Switches in Yeast.Life (Basel, Switzerland) · 2022Review
- Construction of cadmium whole-cell biosensors and circuit amplification.Applied microbiology and biotechnology · 2021Article
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Authors and funding
5 authors.
Funding
Abstract
Directed evolution has been proven as a powerful tool for developing proteins and strains with novel or enhanced features. In this study, a dual selection system was designed to tune the binding specificity of a transcription factor to a particular ligand with the ampicillin resistance gene amp (ON selection) as the positive selection marker and the levansucrase gene sacB (OFF selection) as the negative selection marker. It was applied to the lead responsive transcription factor PbrR in a whole-cell lead biosensor previously constructed in our lab (Jia et al. in Fems Microbiol Lett 365:fny157, 2018). After multiple rounds of ON-OFF selection, two mutants with higher specificity for lead were selected. Structural analysis revealed that the mutation C134 located on the metal-binding loop at the C-terminal of PbrR is likely associated with the enhanced binding to both lead and cadmium. The double mutations D64A and L68S close to the metal-binding residue C79 may lead to the reduced binding specificity toward zinc ions. This dual selection system can be applied to engineer the specificity of other transcription factors and provide fine-tuned tools to synthetic biology.
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