Journal
NUCLEIC ACIDS RESEARCH
Volume 37, Issue 2, Pages -Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkn991
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Funding
- National Institutes of Health [GM077596]
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The assembly of large recombinant DNA encoding a whole biochemical pathway or genome represents a significant challenge. Here, we report a new method, DNA assembler, which allows the assembly of an entire biochemical pathway in a single step via in vivo homologous recombination in Saccharomyces cerevisiae. We show that DNA assembler can rapidly assemble a functional d-xylose utilization pathway (9 kb DNA consisting of three genes), a functional zeaxanthin biosynthesis pathway (11 kb DNA consisting of five genes) and a functional combined d-xylose utilization and zeaxanthin biosynthesis pathway (19 kb consisting of eight genes) with high efficiencies (70100) either on a plasmid or on a yeast chromosome. As this new method only requires simple DNA preparation and one-step yeast transformation, it represents a powerful tool in the construction of biochemical pathways for synthetic biology, metabolic engineering and functional genomics studies.
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