ArticleNucleic acids research2008
Expanded molecular diversity generation during directed evolution by trinucleotide exchange (TriNEx).
Article in Nucleic acids research, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 50 citations in OpenAlex.
- High-throughput methods enabling random duplications, deletions, or nucleotide-constrained mutagenesis of entire DNA motifs.Nucleic acids research · 2026Article
- On the origin of non-specific binders isolated in the selection of phage display peptide libraries.Frontiers in microbiology · 2025Review
- In vitro generation of genetic diversity for directed evolution by error-prone artificial DNA synthesis.Communications biology · 2024Article
- Assessment of Phenotype Relevant Amino Acid Residues in TEM-β-Lactamases by Mathematical Modelling and Experimental Approval.Microorganisms · 2021Article
- Accessing unexplored regions of sequence space in directed enzyme evolution via insertion/deletion mutagenesis.Nature communications · 2020Article
- Functional Trade-Offs in Promiscuous Enzymes Cannot Be Explained by Intrinsic Mutational Robustness of the Native Activity.PLoS genetics · 2016Article
- Structural plasticity of green fluorescent protein to amino acid deletions and fluorescence rescue by folding-enhancing mutations.BMC biochemistry · 2015Article
- Lectin engineering, a molecular evolutionary approach to expanding the lectin utilities.Molecules (Basel, Switzerland) · 2015Review
- Synthetic biology for the directed evolution of protein biocatalysts: navigating sequence space intelligently.Chemical Society reviews · 2015Review
- Directed evolution of GFP with non-natural amino acids identifies residues for augmenting and photoswitching fluorescence.Chemical science · 2015Article
- Article
- Generation of comprehensive transposon insertion mutant library for the model archaeon, Haloferax volcanii, and its use for gene discovery.BMC biology · 2014Article
- An assay to monitor the activity of DNA transposition complexes yields a general quality control measure for transpositional recombination reactions.Mobile genetic elements · 2014Article
- Structural and dynamic changes associated with beneficial engineered single-amino-acid deletion mutations in enhanced green fluorescent protein.Acta crystallographica. Section D, Biological crystallography · 2014Article
- A transposase strategy for creating libraries of circularly permuted proteins.Nucleic acids research · 2012Article
- A method for multi-codon scanning mutagenesis of proteins based on asymmetric transposons.Protein engineering, design & selection : PEDS · 2012Article
- Experimental methods for scanning unnatural amino acid mutagenesis.Methods in molecular biology (Clifton, N.J.) · 2012Article
- PFunkel: efficient, expansive, user-defined mutagenesis.PloS one · 2012Article
- Network models of TEM β-lactamase mutations coevolving under antibiotic selection show modular structure and anticipate evolutionary trajectories.PLoS computational biology · 2011Article
- Applying neutral drift to the directed molecular evolution of a β-glucuronidase into a β-galactosidase: Two different evolutionary pathways lead to the same variant.BMC research notes · 2011Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Trinucleotide exchange (TriNEx) is a method for generating novel molecular diversity during directed evolution by random substitution of one contiguous trinucleotide sequence for another. Single trinucleotide sequences were deleted at random positions in a target gene using the engineered transposon MuDel that were subsequently replaced with a randomized trinucleotide sequence donated by the DNA cassette termed SubSeq(NNN). The bla gene encoding TEM-1 beta-lactamase was used as a model to demonstrate the effectiveness of TriNEx. Sequence analysis revealed that the mutations were distributed throughout bla, with variants containing single, double and triple nucleotide changes. Many of the resulting amino acid substitutions had significant effects on the in vivo activity of TEM-1, including up to a 64-fold increased activity toward ceftazidime and up to an 8-fold increased resistance to the inhibitor clavulanate. Many of the observed amino acid substitutions were only accessible by exchanging at least two nucleotides per codon, including charge-switch (R164D) and aromatic substitution (W165Y) mutations. TriNEx can therefore generate a diverse range of protein variants with altered properties by combining the power of site-directed saturation mutagenesis with the capacity of whole-gene mutagenesis to randomly introduce mutations throughout a 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.