ArticleACS synthetic biology2012
Multiplexed in vivo His-tagging of enzyme pathways for in vitro single-pot multienzyme catalysis.
Article in ACS synthetic biology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers.
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Who cites it
46 citing papers in PubMed.
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Coselection of BAC for Escherichia coli chromosomal DNA multiplex automated genome engineering.Biotechnology letters · 2024Article
- The Long Road to a Synthetic Self-Replicating Central Dogma.Biochemistry · 2023Review
- On-demand biomanufacturing through synthetic biology approach.Materials today. Bio · 2023Article
- Cell-Free Display Techniques for Protein Evolution.Advances in biochemical engineering/biotechnology · 2023Article
- Engineering consortia by polymeric microbial swarmbots.Nature communications · 2022Article
- Self-assembled nanoparticle-enzyme aggregates enhance functional protein production in pure transcription-translation systems.PloS one · 2022Article
- Cell-Free PURE System: Evolution and Achievements.Biodesign research · 2022Review
- Ribosome purification fromBiotechnology notes (Amsterdam, Netherlands) · 2022Article
- A lysate proteome engineering strategy for enhancing cell-free metabolite production.Metabolic engineering communications · 2021Article
- Recombineering and MAGE.Nature reviews. Methods primers · 2021Article
- A partially self-regenerating synthetic cell.Nature communications · 2020Article
- In vitro self-replication and multicistronic expression of large synthetic genomes.Nature communications · 2020Article
- Review
- Emerging Methods for Efficient and Extensive Incorporation of Non-canonical Amino Acids Using Cell-Free Systems.Frontiers in bioengineering and biotechnology · 2020Review
- Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology.Frontiers in bioengineering and biotechnology · 2020Review
- Multiplex transcriptional characterizations across diverse bacterial species using cell-free systems.Molecular systems biology · 2019Article
- A versatile platform strain for high-fidelity multiplex genome editing.Nucleic acids research · 2019Article
- Exploring the Potential of Cell-Free Protein Synthesis for Extending the Abilities of Biological Systems.Frontiers in bioengineering and biotechnology · 2019Review
- AnSynthetic and systems biotechnology · 2018Review
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Authors and funding
10 authors.
Funding
Abstract
Protein pathways are dynamic and highly coordinated spatially and temporally, capable of performing a diverse range of complex chemistries and enzymatic reactions with precision and at high efficiency. Biotechnology aims to harvest these natural systems to construct more advanced in vitro reactions, capable of new chemistries and operating at high yield. Here, we present an efficient Multiplex Automated Genome Engineering (MAGE) strategy to simultaneously modify and co-purify large protein complexes and pathways from the model organism Escherichia coli to reconstitute functional synthetic proteomes in vitro. By application of over 110 MAGE cycles, we successfully inserted hexa-histidine sequences into 38 essential genes in vivo that encode for the entire translation machinery. Streamlined co-purification and reconstitution of the translation protein complex enabled protein synthesis in vitro. Our approach can be applied to a growing area of applications in in vitro one-pot multienzyme catalysis (MEC) to manipulate or enhance in vitro pathways such as natural product or carbohydrate biosynthesis.
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Registered trials
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