ArticleMethods in molecular biology (Clifton, N.J.)2013
Construction and engineering of large biochemical pathways via DNA assembler.
Article in Methods in molecular biology (Clifton, N.J.), 2013. 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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Who cites it
11 citing papers in PubMed.
- How many plasmids can bacteria carry? A synthetic biology perspective.Open biology · 2025Review
- Heterologous overproduction of oviedomycin by refactoring biosynthetic gene cluster and metabolic engineering of host strain Streptomyces coelicolor.Microbial cell factories · 2023Article
- Easy efficient HDR-based targeted knock-in inBioengineered · 2022Article
- Review
- Genomic Considerations for the Modification ofMicroorganisms · 2020Review
- Genetic platforms for heterologous expression of microbial natural products.Natural product reports · 2019Review
- Heterologous expression-facilitated natural products' discovery in actinomycetes.Journal of industrial microbiology & biotechnology · 2019Review
- Characterization of a panARS-based episomal vector in the methylotrophic yeast Pichia pastoris for recombinant protein production and synthetic biology applications.Microbial cell factories · 2016Article
- Development of a modularized two-step (M2S) chromosome integration technique for integration of multiple transcription units inBiotechnology for biofuels · 2016Article
- Robust orthogonal recombination system for versatile genomic elements rearrangement in yeast Saccharomyces cerevisiae.Scientific reports · 2015Article
- The evolving art of creating genetic diversity: From directed evolution to synthetic biology.Biotechnology advancesReview
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2 authors.
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Abstract
DNA assembler enables rapid construction and engineering of biochemical pathways in a one-step fashion by exploitation of the in vivo homologous recombination mechanism in Saccharomyces cerevisiae. It has many applications in pathway engineering, metabolic engineering, combinatorial biology, and synthetic biology. Here we use two examples including the zeaxanthin biosynthetic pathway and the aureothin biosynthetic gene cluster to describe the key steps in the construction of pathways containing multiple genes using the DNA assembler approach. Methods for construct design, pathway assembly, pathway confirmation, and functional analysis are shown. The protocol for fine genetic modifications such as site-directed mutagenesis for engineering the aureothin gene cluster is also illustrated.
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