ArticleCell reports methods2023
Engineering E. coli strains using antibiotic-resistance-gene-free plasmids.
Article in Cell reports methods, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
- Gut microbial signatures in poultry under low-protein diets and lysine restriction: implications for precision nutrition.Veterinary and animal science · 2026Review
- Engineering rhizobacterial communities for soil and plant health.Microbiology (Reading, England) · 2026Review
- Resilience of recombinant antibiotic resistance gene-containing plasmids against common cell culture disposal methods.Cell reports methods · 2026Article
- Hydrogel platforms for engineered live biotherapeutics: materials, microbial integration and clinical potential.RSC pharmaceutics · 2026Review
- tRNA-Mediated Plasmid Stabilization for Antibiotic-Free Applications inACS synthetic biology · 2026Article
- Probiotic-Based Materials as Living Therapeutics.Advanced materials (Deerfield Beach, Fla.) · 2026Review
- On the Choice of the Right Plasmid Vector(s) in the Times of Synthetic Biology.Microbial biotechnology · 2025Review
- How many plasmids can bacteria carry? A synthetic biology perspective.Open biology · 2025Review
- Be a GEM: Biocontained, environmentally applied, genetically engineered microbes.Advanced drug delivery reviews · 2025Review
- Prevalence and molecular characterization of multi-drug and extreme drug resistant Escherichia coli in companion animals in Bangladesh.Scientific reports · 2025Article
- Knockout of the Agrobacterium ILVC gene generates a valine-isoleucine auxotrophic strain for plant transformation.Transgenic research · 2025Article
- Precision engineering of the probioticApplied and environmental microbiology · 2025Article
- A mathematical model of cell-free transcription-translation with plasmid crosstalk.Synthetic biology (Oxford, England) · 2025Article
- One-for-all gene inactivation via PAM-independent base editing in bacteria.The Journal of biological chemistry · 2025Article
- Influence of Environmental Conditions on the Escape Rates of Biocontained Genetically Engineered Microbes.Environmental science & technology · 2024Article
- Dynamic control of the plasmid copy number maintained without antibiotics in Escherichia coli.Journal of biological engineering · 2024Article
- Earth: Extinguishing anthropogenic risks through harmonization.New biotechnology · 2024Review
- Multiplex genetic manipulations in Clostridium butyricum and Clostridium sporogenes to secrete recombinant antigen proteins for oral-spore vaccination.Microbial cell factories · 2024Article
Corrections and comments
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Authors and funding
7 authors.
Funding
Abstract
We created a generalizable pipeline for antibiotic-resistance-gene-free plasmid (ARGFP)-based cloning using a dual auxotrophic- and essential-gene-based selection strategy. We use auxotrophic selection to construct plasmids in engineered E. coli DH10B cloning strains and both auxotrophic- and essential-gene-based selection to (1) select for recombinant strains and (2) maintain a plasmid in E. coli Nissle 1917, a common chassis for engineered probiotic applications, and E. coli MG1655, the laboratory "wild-type" E. coli strain. We show that our approach has comparable efficiency to that of antibiotic-resistance-gene-based cloning. We also show that the double-knockout Nissle and MG1655 strains are simple to transform with plasmids of interest. Notably, we show that the engineered Nissle strains are amenable to long-term plasmid maintenance in repeated culturing as well as in the mouse gut, demonstrating the potential for broad applications while minimizing the risk of antibiotic resistance spread via horizontal gene transfer.
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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.