ArticleJournal of the American Chemical Society2005
Polymerase evolution: efforts toward expansion of the genetic code.
Article in Journal of the American Chemical Society, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
14 citing papers in PubMed, 64 citations in OpenAlex.
- DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions.International journal of molecular sciences · 2023Review
- From polymerase engineering to semi-synthetic life: artificial expansion of the central dogma.RSC chemical biology · 2022Review
- Article
- Engineering Polymerases for New Functions.Trends in biotechnology · 2019Review
- Exploring the Chemistry of Genetic Information Storage and Propagation through Polymerase Engineering.Accounts of chemical research · 2017Review
- Modified Nucleoside Triphosphates for In-vitro Selection Techniques.Frontiers in chemistry · 2016Review
- DNA sequencing using polymerase substrate-binding kinetics.Nature communications · 2015Article
- Directed polymerase evolution.FEBS letters · 2014Review
- DNA polymerases engineered by directed evolution to incorporate non-standard nucleotides.Frontiers in microbiology · 2014Review
- A general strategy for the evolution of bond-forming enzymes using yeast display.Proceedings of the National Academy of Sciences of the United States of America · 2011Article
- Directed evolution of DNA polymerases for next-generation sequencing.Angewandte Chemie (International ed. in English) · 2010Article
- Evolving a polymerase for hydrophobic base analogues.Journal of the American Chemical Society · 2009Article
- Optimization of an unnatural base pair toward natural-like replication.Journal of the American Chemical Society · 2009Article
- Discovery, characterization, and optimization of an unnatural base pair for expansion of the genetic alphabet.Journal of the American Chemical Society · 2008Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Genetic information is encoded by, but potentially not limited to, a four-letter alphabet. A variety of predominantly hydrophobic nucleobase analogues that form self-pairs in DNA have been examined as third base pair candidates. For example, the PICS self-pair is both stable in duplex DNA and synthesized by some wild-type polymerases with reasonable efficiency. These efforts to expand the genetic code are expected to be facilitated by optimizing both the unnatural nucleobase analogues and the polymerases that replicate them. Here, we report the use of an activity-based selection system to evolve a DNA polymerase that more efficiently replicates DNA containing the PICS self-pair. The selection system is based on the co-display on phage of DNA polymerase libraries and a DNA substrate containing the self-pair. Only polymerases that accept the unnatural substrate incorporate a biotin-dUTP to the attached primer and may then be isolated on a streptavidin solid support. A mutant of Sf polymerase, P2, was evolved which both inserts dPICSTP opposite dPICS in the template and extends the unnatural primer terminus by incorporation of the next correct natural dNTP, where the parental enzyme catalyzes neither step at detectable rates. P2 was found to be a triple mutant of Sf, with the mutations F598I, I614F, and Q489H. The evolved properties of P2, as well as the observed mutations, are consistent with an increased affinity for the DNA primer-template containing the self-pair.
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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.