ArticlePloS one2015
An Integrated System for Precise Genome Modification in Escherichia coli.
Article in PloS one, 2015. 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, 37 citations in OpenAlex.
- A comprehensive review of genomic-scale genetic engineering as a strategy to improve bacterial productivity.Microbiology (Reading, England) · 2025Review
- The combination of active partitioning and toxin-antitoxin systems is most advantageous for low-copy plasmid fitness.Nature communications · 2025Article
- Engineered Escherichia coli for the in situ secretion of therapeutic nanobodies in the gut.Cell host & microbe · 2023Article
- SEVA 4.0: an update of the Standard European Vector Architecture database for advanced analysis and programming of bacterial phenotypes.Nucleic acids research · 2023Article
- Genetic components ofFrontiers in microbiology · 2023Article
- Targeted insertion of large genetic payloads using cas directed LINE-1 reverse transcriptase.Scientific reports · 2021Article
- Biosynthesis of l-phosphinothricin with enzymes from chromosomal integrated expression in3 Biotech · 2021Article
- NgAgo possesses guided DNA nicking activity.Nucleic acids research · 2021Article
- Genome Editing in Bacteria: CRISPR-Cas and Beyond.Microorganisms · 2021Review
- Article
- Bacterial Genetic Engineering by Means of Recombineering for Reverse Genetics.Frontiers in microbiology · 2020Review
- Testing the retroelement invasion hypothesis for the emergence of the ancestral eukaryotic cell.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
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- Article
- Oligo- and dsDNA-mediated genome editing using a tetA dual selection system in Escherichia coli.PloS one · 2017Article
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- Real-time transposable element activity in individual live cells.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe an optimized system for the easy, effective, and precise modification of the Escherichia coli genome. Genome changes are introduced first through the integration of a 1.3 kbp Landing Pad consisting of a gene conferring resistance to tetracycline (tetA) or the ability to metabolize the sugar galactose (galK). The Landing Pad is then excised as a result of double-strand breaks by the homing endonuclease I-SceI, and replaced with DNA fragments bearing the desired change via λ-Red mediated homologous recombination. Repair of the double strand breaks and counterselection against the Landing Pad (using NiCl2 for tetA or 2-deoxy-galactose for galK) allows the isolation of modified bacteria without the use of additional antibiotic selection. We demonstrate the power of this method to make a variety of genome modifications: the exact integration, without any extraneous sequence, of the lac operon (~6.5 kbp) to any desired location in the genome and without the integration of antibiotic markers; the scarless deletion of ribosomal rrn operons (~6 kbp) through either intrachromosomal or oligonucleotide recombination; and the in situ fusion of native genes to fluorescent reporter genes without additional perturbation.
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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.