ArticleNucleic acids research2007
Selection of restriction endonucleases using artificial cells.
Article in Nucleic acids research, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 30 citations in OpenAlex.
- A sequential one-pot approach for rapid and convenient characterization of putative restriction-modification systems.mSystems · 2023Article
- Split & mix assembly of DNA libraries for ultrahigh throughput on-bead screening of functional proteins.Nucleic acids research · 2020Article
- Natural selection in compartmentalized environment with reshuffling.Journal of mathematical biology · 2019Article
- High-throughput screening of biomolecules using cell-free gene expression systems.Synthetic biology (Oxford, England) · 2018Review
- Engineering of customized meganucleases via in vitro compartmentalization and in cellulo optimization.Methods in molecular biology (Clifton, N.J.) · 2015Article
- High-throughput bead-based identification of structure-switching aptamer beacons.Chembiochem : a European journal of chemical biology · 2014Article
- Redesign of extensive protein-DNA interfaces of meganucleases using iterative cycles of in vitro compartmentalization.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
- Piecing Together Cell-like Systems.Current organic chemistry · 2013Article
- A Concept for Selection of Codon-Suppressor tRNAs Based on Read-Through Ribosome Display in an In Vitro Compartmentalized Cell-Free Translation System.Journal of nucleic acids · 2012Article
- In vitro genetic reconstruction of bacterial transcription initiation by coupled synthesis and detection of RNA polymerase holoenzyme.Nucleic acids research · 2010Article
- Selection of bacteriophage lambda integrases with altered recombination specificity by in vitro compartmentalization.Nucleic acids research · 2010Article
- A novel sequence-specific RNA quantification method using nicking endonuclease, dual-labeled fluorescent DNA probe, and conformation-interchangeable oligo-DNA.RNA (New York, N.Y.) · 2008Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe in this article an in vitro system for the selection of restriction endonucleases using artificial cells. The artificial cells are generated in the form of a water-in-oil emulsion by in vitro compartmentalization. Each aqueous compartment contains a reconstituted transcription/translation mix along with the dispersed DNA templates. In the compartments containing endonuclease genes, an endonuclease expressed in vitro cleaves its own DNA template adjacent to the gene, leaving a sticky end. The pooled DNA templates are then ligated to an adaptor with a compatible end. The endonuclease genes are then enriched by adaptor-specific PCR on the ligation mix. We demonstrate that the system can achieve at least 100-fold enrichment in a single round of selection. It is sensitive enough to enrich an active endonuclease gene from a 1:10(5) model library in 2-3 rounds of selection. Finally, we describe experiments where we selected endonuclease genes directly from a bacterial genomic DNA source in three rounds of selections: the known PstI gene from Providencia stuartii and the new TspMI gene from Thermus sp. manalii. This method provides a unique tool for cloning restriction endonuclease genes and has many other potential applications.
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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.