ArticleNucleic acids research1996
Hypermutagenic PCR involving all four transitions and a sizeable proportion of transversions.
Article in Nucleic acids research, 1996. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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Who cites it
40 citing papers in PubMed, 143 citations in OpenAlex.
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- Tuning Thermostability and Catalytic Efficiency of Aflatoxin-Degrading Enzyme by Error-prone PCR.Applied microbiology and biotechnology · 2023Article
- Engineered Recombinant PON1-OPH Fusion Hybrids: Potentially Effective Catalytic Bioscavengers against Organophosphorus Nerve Agent Analogs.Journal of microbiology and biotechnology · 2021Article
- Fast reversibly photoswitching red fluorescent proteins for live-cell RESOLFT nanoscopy.Nature methods · 2018Article
- Targeted mutagenesis: A sniper-like diversity generator in microbial engineering.Synthetic and systems biotechnology · 2017Review
- Biosynthesis of selenate reductase in Salmonella enterica: critical roles for the signal peptide and DmsD.Microbiology (Reading, England) · 2016Article
- KillerOrange, a Genetically Encoded Photosensitizer Activated by Blue and Green Light.PloS one · 2015Article
- DNA polymerases engineered by directed evolution to incorporate non-standard nucleotides.Frontiers in microbiology · 2014Review
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- Distant and new mutations in CTX-M-1 beta-lactamase affect cefotaxime hydrolysis.Antimicrobial agents and chemotherapy · 2011Article
- Deep sequencing analysis of mutations resulting from the incorporation of dNTP analogs.Nucleic acids research · 2010Article
- All-codon scanning identifies p53 cancer rescue mutations.Nucleic acids research · 2010Article
- Selection of bacteriophage lambda integrases with altered recombination specificity by in vitro compartmentalization.Nucleic acids research · 2010Article
- Enhancing the prebiotic relevance of a set of covalently self-assembling, autorecombining RNAs through in vitro selection.Journal of molecular evolution · 2010Article
- TtsI regulates symbiotic genes in Rhizobium species NGR234 by binding to tts boxes.Molecular microbiology · 2008Article
- Darwinian evolution on a chip.PLoS biology · 2008Article
- Emergence of a fast-reacting ribozyme that is capable of undergoing continuous evolution.Proceedings of the National Academy of Sciences of the United States of America · 2007Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Very complex mutant libraries of the dihydrofolate reductase (DHFR) gene encoded by the Escherichia coli plasmid R67 were created using hypermutagenic PCR with biased deoxynucleotide triphosphate (dNTP) concentrations. Exploiting the particular stability of the G:T mismatch, the DHFR gene could be enriched in A+T by employing biased deoxypyrimidine triphosphate concentrations, i.e. [dTTP] > [dCTP]. A sizeable fraction of hypermutants were functional. A combination of [dTTP] > [dCTP] and [dGTP] > [dATP] biases generated mutations at unexpectedly low frequencies. This could be overcome by the addition of Mn2+ cations. Overall mutation frequencies of 10% per amplification (range 4-18% per clone) could be attained. All four transitions and a smaller number of transversions were produced throughout the gene. PCR mutagenesis could be so extensive as to inactivate all amplified versions of the gene.
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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.