DNA assembler, an in vivo genetic method for rapid construction of biochemical pathways

Zengyi Shao, Hua Zhao, Huimin Zhao

Research output: Contribution to journalArticlepeer-review

Abstract

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 (70 - 100%) 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.

Original languageEnglish (US)
Article numbere16
JournalNucleic acids research
Volume37
Issue number2
DOIs
StatePublished - 2009

ASJC Scopus subject areas

  • Genetics

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