ArticleBioengineered bugs
A place for everything: chromosomal integration of large constructs.
Article in Bioengineered bugs. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 28 citations in OpenAlex.
- 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
- Article
- Real-time transposable element activity in individual live cells.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- Article
- Automated single cell microbioreactor for monitoring intracellular dynamics and cell growth in free solution.Lab on a chip · 2014Article
- High-efficiency scarless genetic modification in Escherichia coli by using lambda red recombination and I-SceI cleavage.Applied and environmental microbiology · 2014Article
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Authors and funding
2 authors at 1 institution in 1 country.
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Abstract
We have developed an easy, reliable two-step method for the insertion of large DNA fragments into any desired location in the E. coli chromosome. The method is based on the recombineering of a small (∼1.3 kbp) "Landing Pad" into the chromosome at the insertion site, to which the large construct is subsequently delivered via I-SceI endonuclease excision from a donor plasmid. To demonstrate the power of this method, we here show the insertion of a fragment containing the entire lac operon (∼9 kbp) into four predefined novel locations in the E. coli chromosome, a feat not possible with existing technologies. In addition, the chromosomal breaks induced by landing pad excision provide sufficient selective pressure that positive selection by antibiotics is unnecessary, making precise, exact insertion without extraneous sequence possible.
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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.