ArticleScientific reports2019
Golden Mutagenesis: An efficient multi-site-saturation mutagenesis approach by Golden Gate cloning with automated primer design.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- Artificial intelligence and automation in enzyme engineering: evolution, advances, and future perspectives.Bioresources and bioprocessing · 2026Review
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- Reverse engineering of the pattern recognition receptor FLS2 reveals key design principles of broader recognition spectra against evading flg22 epitopes.Nature plants · 2025Article
- Biosynthesis and bioactivity of anti-inflammatory triterpenoids in Calendula officinalis.Nature communications · 2025Article
- Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesis.Nature communications · 2025Article
- Engineering of Conserved Sequence Motif 1 Residues in Halohydrin Dehalogenase HheC Simultaneously Enhances Activity, Stability, and Enantioselectivity.ACS catalysis · 2025Article
- Generating Site Saturation Mutagenesis Libraries and Transferring Them to Broad Host-Range Plasmids Using Golden Gate Cloning.Methods in molecular biology (Clifton, N.J.) · 2025Article
- GGAssembler: Precise and economical design and synthesis of combinatorial mutation libraries.Protein science : a publication of the Protein Society · 2024Article
- Exploring class III cellobiose dehydrogenase: sequence analysis and optimized recombinant expression.Microbial cell factories · 2024Article
- Intein-based thermoregulated meganucleases for containment of genetic material.Nucleic acids research · 2024Article
- Enzymatic Routes for Chiral Amine Synthesis: Protein Engineering and Process Optimization.Biologics : targets & therapy · 2024Review
- Standardizing cassette-based deep mutagenesis by Golden Gate assembly.Biotechnology and bioengineering · 2024Article
- Large-scale, in-cell photocrosslinking at single-residue resolution reveals the molecular basis for glucocorticoid receptor regulation by immunophilins.Nature structural & molecular biology · 2023Article
- Amino acid positions near the active site determine the reduced activity of human ACOD1 compared to murine ACOD1.Scientific reports · 2023Article
- Ultradeep characterisation of translational sequence determinants refutes rare-codon hypothesis and unveils quadruplet base pairing of initiator tRNA and transcript.Nucleic acids research · 2023Article
- A growth selection system for the directed evolution of amine-forming or converting enzymes.Nature communications · 2022Article
- A multifunctional system for genome editing and large-scale interspecies gene transfer.Nature communications · 2022Article
- Making the cut with protease engineering.Cell chemical biology · 2022Review
- Versatile selective evolutionary pressure using synthetic defect in universal metabolism.Nature communications · 2021Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Site-directed methods for the generation of genetic diversity are essential tools in the field of directed enzyme evolution. The Golden Gate cloning technique has been proven to be an efficient tool for a variety of cloning setups. The utilization of restriction enzymes which cut outside of their recognition domain allows the assembly of multiple gene fragments obtained by PCR amplification without altering the open reading frame of the reconstituted gene. We have developed a protocol, termed Golden Mutagenesis that allows the rapid, straightforward, reliable and inexpensive construction of mutagenesis libraries. One to five amino acid positions within a coding sequence could be altered simultaneously using a protocol which can be performed within one day. To facilitate the implementation of this technique, a software library and web application for automated primer design and for the graphical evaluation of the randomization success based on the sequencing results was developed. This allows facile primer design and application of Golden Mutagenesis also for laboratories, which are not specialized in molecular biology.
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