ArticleMicrobial biotechnology2018
A standardized workflow for surveying recombinases expands bacterial genome-editing capabilities.
Article in Microbial biotechnology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
- Synthetic Biology of Plants and Microbes for Agriculture, Environment, and Future Applications.Chemical reviews · 2026Review
- Alternate routes to acetate tolerance lead to varied isoprenol production from mixed carbon sources inApplied and environmental microbiology · 2025Article
- Metagenomics harvested genus-specific single-stranded DNA-annealing proteins improve and expand recombineering in Pseudomonas species.Nucleic acids research · 2023Article
- An expanded CRISPR-Cas9-assisted recombineering toolkit for engineering genetically intractable Pseudomonas aeruginosa isolates.Nature protocols · 2023Review
- Precise genome engineering in Pseudomonas using phage-encoded homologous recombination and the Cascade-Cas3 system.Nature protocols · 2023Review
- SURE editing: combining oligo-recombineering and programmable insertion/deletion of selection markers to efficiently edit the Mycoplasma pneumoniae genome.Nucleic acids research · 2022Article
- Recombineering in Non-Model Bacteria.Current protocols · 2022Article
- Evolution of biofilm-adapted gene expression profiles in lasR-deficient clinical Pseudomonas aeruginosa isolates.NPJ biofilms and microbiomes · 2022Article
- Propagation of Recombinant Genes through Complex Microbiomes with Synthetic Mini-RP4 Plasmid Vectors.Biodesign research · 2022Article
- Genomic characterization of a new phage BUCT541 againstFrontiers in microbiology · 2022Article
- ReScribe: An Unrestrained Tool Combining Multiplex Recombineering and Minimal-PAM ScCas9 for Genome RecodingACS synthetic biology · 2021Article
- Recombineering and MAGE.Nature reviews. Methods primers · 2021Article
- Article
- Improved bacterial recombineering by parallelized protein discovery.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- High-Efficiency Multi-site Genomic Editing of Pseudomonas putida through Thermoinducible ssDNA Recombineering.iScience · 2020Article
- Rapid Evolution of Reduced Susceptibility against a Balanced Dual-Targeting Antibiotic through Stepping-Stone Mutations.Antimicrobial agents and chemotherapy · 2019Article
- Single-Stranded DNA-Binding Protein and Exogenous RecBCD Inhibitors Enhance Phage-Derived Homologous Recombination in Pseudomonas.iScience · 2019Article
- Evolutionary highways to persistent bacterial infection.Nature communications · 2019Article
- Directed evolution of multiple genomic loci allows the prediction of antibiotic resistance.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
- Phage Therapy: What Have We Learned?Viruses · 2018Review
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
Bacterial recombineering typically relies on genomic incorporation of synthetic oligonucleotides as mediated by Escherichia coli λ phage recombinase β - an occurrence largely limited to enterobacterial strains. While a handful of similar recombinases have been documented, recombineering efficiencies usually fall short of expectations for practical use. In this work, we aimed to find an efficient Recβ homologue demonstrating activity in model soil bacterium Pseudomonas putida EM42. To this end, a genus-wide protein survey was conducted to identify putative recombinase candidates for study. Selected novel proteins were assayed in a standardized test to reveal their ability to introduce the K43T substitution into the rpsL gene of P. putida. An ERF superfamily protein, here termed Rec2, exhibited activity eightfold greater than that of the previous leading recombinase. To bolster these results, we demonstrated Rec2 ability to enter a range of mutations into the pyrF gene of P. putida at similar frequencies. Our results not only confirm the utility of Rec2 as a Recβ functional analogue within the P. putida model system, but also set a complete workflow for deploying recombineering in other bacterial strains/species. Implications range from genome editing of P. putida for metabolic engineering to extended applications within other Pseudomonads - and beyond.
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