ArticleJournal of biological engineering2015
A novel approach for metabolic pathway optimization: Oligo-linker mediated assembly (OLMA) method.
Article in Journal of biological engineering, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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11 citing papers in PubMed.
- Inducible CRISPRi enables efficient and high-fidelity genome editing in Streptomyces.Nucleic acids research · 2026Article
- Fine-Tuning Genetic Circuits via Host Context and RBS Modulation.ACS synthetic biology · 2025Article
- A flexible, modular and versatile functional part assembly toolkit for gene cluster engineering inSynthetic and systems biotechnology · 2024Article
- Insights into the synthesis, engineering, and functions of microbial pigments inFrontiers in microbiology · 2024Review
- Metabolic engineering of Escherichia coli for high-level production of violaxanthin.Microbial cell factories · 2023Article
- Optimizing the downstream MVA pathway using a combination optimization strategy to increase lycopene yield in Escherichia coli.Microbial cell factories · 2022Article
- Multiplexed Promoter Engineering for Improving Thaxtomin A Production in HeterologousLife (Basel, Switzerland) · 2022Article
- Application of combinatorial optimization strategies in synthetic biology.Nature communications · 2020Review
- Design and tailoring of an artificial DNA scaffolding system for efficient lycopene synthesis using zinc-finger-guided assembly.Journal of industrial microbiology & biotechnology · 2020Article
- Analysis and expression of the carotenoid biosynthesis genes from Deinococcus wulumuqiensis R12 in engineered Escherichia coli.AMB Express · 2018Article
- High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment.Scientific reports · 2017Article
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4 authors.
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Abstract
backgroundImbalances in gene expression of a metabolic pathway can result in less-yield of the desired products. Several targets were intensively investigated to balance the gene expression, such as promoter, ribosome binding site (RBS), the order of genes, as well as the species of the enzymes. However, the capability of simultaneous manipulation of multiple targets still needs to be explored.
resultsWe reported a new DNA assembling method to vary all the above types of regulatory targets simultaneously, named oligo-linker mediated assembly (OLMA) method, which can incorporate up to 8 targets in a single assembly step. Two experimental cases were used to demonstrate the capability of the method: (1) assembly of multiple pieces of lacZ expression cassette; (2) optimization of four enzymes in lycopene biosynthetic pathway. Our results indicated that the OLMA method not only exploited larger combinatorial space, but also reduced the inefficient mutants.
conclusionsThe unique feature of oligo-linker mediated assembly (OLMA) method is inclusion of a set of chemically synthetic double-stranded DNA oligo library, which can be designed as promoters and RBSs, or designed with different overhang to bridge the genes in different orders. The inclusion of the oligos resulted in a PCR-free and zipcode-free DNA assembly reaction for OLMA.
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