ArticleJournal of industrial microbiology & biotechnology2018
Genetic tools for reliable gene expression and recombineering in Pseudomonas putida.
Article in Journal of industrial microbiology & biotechnology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 74 papers.
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Who cites it
74 citing papers in PubMed.
- Cascading recombinase memory switch for programmable and stable gene expression in Pseudomonas putida.Nucleic acids research · 2026Article
- LysR-type transcriptional regulator TtdR regulates both ethylene glycol and polyhydroxyalkanoate (PHA) metabolism in Pseudomonas umsongensis GO16.Applied microbiology and biotechnology · 2026Article
- Rhodo-Box: A Synthetic Biology Toolbox to Facilitate Metabolic Engineering ofACS synthetic biology · 2026Article
- Biochemical Upcycling of PET via Glycolysis and Engineered Microbial Consortia.Microbial biotechnology · 2026Article
- Microgravity effects on secondary metabolism of plant-affecting bacteria.Microbiology spectrum · 2026Article
- Synthetic Biology of Plants and Microbes for Agriculture, Environment, and Future Applications.Chemical reviews · 2026Review
- CRISPR/Cas9-Mediated Metabolic Engineering of EndophyticACS omega · 2026Article
- Implementation of the synthetic arabinose 5-phosphate dependent glycolaldehyde assimilation (SAGA) pathway inFrontiers in bioengineering and biotechnology · 2026Article
- Genetically Reprogramming Crops and Rhizobacteria for Nutritional Iron Biofortification.ACS synthetic biology · 2025Review
- The link of carbon catabolite repression elements, small RNAs CrcY and CrcZ and polyhydroxyalkanoate metabolism in Pseudomonas putida KT2440.Biotechnology for biofuels and bioproducts · 2025Article
- Binary vector copy number engineering improves Agrobacterium-mediated transformation.Nature biotechnology · 2025Article
- Protein-Protein Complex Stability Controls Substrate Scope in a β-Ketoacyl-ACP Reductase Specific for Medium Chains.Angewandte Chemie (International ed. in English) · 2025Article
- Lignin-Degrading Enzymes and the Potential ofMicroorganisms · 2025Review
- Rational Design Assisted by Evolutionary Engineering Allows (De)Construction and Optimization of Complex Phenotypes in Pseudomonas putida KT2440.Microbial biotechnology · 2025Article
- Engineering a phi15-based expression system for stringent gene expression in Pseudomonas putida.Communications biology · 2025Article
- Fine-Tuning Genetic Circuits via Host Context and RBS Modulation.ACS synthetic biology · 2025Article
- A Novel Membrane-Associated Protein Aids Bacterial Colonization of Maize.ACS synthetic biology · 2025Article
- Addressing genome scale design tradeoffs in Pseudomonas putida for bioconversion of an aromatic carbon source.NPJ systems biology and applications · 2025Article
- A single-plasmid-based, easily curable CRISPR/Cas9 system for rapid, iterative genome editing in Pseudomonas putida KT2440.Microbial cell factories · 2024Article
- Phenotypic Plasticity During Organofluorine Degradation Revealed by Adaptive Evolution.Microbial biotechnology · 2024Article
14 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors.
Funding
Abstract
Pseudomonas putida is a promising bacterial host for producing natural products, such as polyketides and nonribosomal peptides. In these types of projects, researchers need a genetic toolbox consisting of plasmids, characterized promoters, and techniques for rapidly editing the genome. Past reports described constitutive promoter libraries, a suite of broad host range plasmids that replicate in P. putida, and genome-editing methods. To augment those tools, we have characterized a set of inducible promoters and discovered that IPTG-inducible promoter systems have poor dynamic range due to overexpression of the LacI repressor. By replacing the promoter driving lacI expression with weaker promoters, we increased the fold induction of an IPTG-inducible promoter in P. putida KT2440 to 80-fold. Upon discovering that gene expression from a plasmid was unpredictable when using a high-copy mutant of the BBR1 origin, we determined the copy numbers of several broad host range origins and found that plasmid copy numbers are significantly higher in P. putida KT2440 than in the synthetic biology workhorse, Escherichia coli. Lastly, we developed a λRed/Cas9 recombineering method in P. putida KT2440 using the genetic tools that we characterized. This method enabled the creation of scarless mutations without the need for performing classic two-step integration and marker removal protocols that depend on selection and counterselection genes. With the method, we generated four scarless deletions, three of which we were unable to create using a previously established genome-editing technique.
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