ArticleBiotechnology for biofuels2019
A seamless and iterative DNA assembly method named PS-Brick and its assisted metabolic engineering for threonine and 1-propanol production.
Article in Biotechnology for biofuels, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed, 18 citations in OpenAlex.
- Technologies for engineering repetitive DNA.Quantitative biology (Beijing, China) · 2026Review
- UniClo technology exploits methylation for universal scarless DNA assembly.Biology methods & protocols · 2026Review
- Genetic make-up and regulation of the L-lysine biosynthesis pathway inMicrobial cell (Graz, Austria) · 2026Article
- Propanol production through microbial fermentation of biomass.Bioresources and bioprocessing · 2025Review
- Development of a lysine biosensor for the dynamic regulation of cadaverine biosynthesis in E. coli.Microbial cell factories · 2025Article
- UniClo: scarless hierarchical DNA assembly without sequence constraint.Nucleic acids research · 2025Article
- Engineering the hyperthermophilic archaeonApplied and environmental microbiology · 2025Article
- Recent Advances in Metabolic Engineering for the Biosynthesis of Phosphoenol Pyruvate-Oxaloacetate-Pyruvate-Derived Amino Acids.Molecules (Basel, Switzerland) · 2024Review
- Multivariate modular metabolic engineering for enhanced L-methionine biosynthesis in Escherichia coli.Biotechnology for biofuels and bioproducts · 2023Article
- Development and Application of CRISPR/Cas in Microbial Biotechnology.Frontiers in bioengineering and biotechnology · 2020Review
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundDNA assembly is an essential technique enabling metabolic engineering and synthetic biology. Combining novel DNA assembly technologies with rational metabolic engineering can facilitate the construction of microbial cell factories. Amino acids and derived biochemicals are important products in industrial biotechnology with wide application and huge markets. DNA assembly scenarios encountered in metabolic engineering for the construction of amino acid and related compound producers, such as design-build-test-learn cycles, construction of precise genetic circuits and repetitive DNA molecules, usually require for iterative, scarless and repetitive sequence assembly methods, respectively.
resultsRestriction endonuclease (RE)-assisted strategies constitute one of the major categories of DNA assembly. Here, we developed a Type IIP and IIS RE-assisted method named PS-Brick that comprehensively takes advantage of the properties of PCR fragments and REs for iterative, seamless and repetitive sequence assembly. One round of PS-Brick reaction using purified plasmids and PCR fragments was accomplished within several hours, and transformation of the resultant reaction product from this PS-Brick assembly reaction exhibited high efficiency (10
conclusionsTo the best of our knowledge, the PS-Brick framework is the first RE-assisted DNA assembly method using the strengths of both Type IIP and IIS REs. In this study, PS-Brick was demonstrated to be an efficient DNA assembly method for pathway construction and genome editing and was successfully applied in design-build-test-learn (DBTL) cycles of metabolic engineering for the production of threonine and threonine-derived 1-propanol. The PS-Brick presents a valuable addition to the current toolbox of synthetic biology and metabolic engineering.
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