articleNucleic Acids ResearchDec 12, 2008GOLD OA

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

ZSZengyi ShaoHZHua ZhaoHZHuimin Zhao

University of Illinois Urbana-Champaign

PubMed
Indexed incrossrefdoajpubmed

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 (approximately 9 kb DNA consisting of three genes), a functional zeaxanthin biosynthesis pathway (approximately 11 kb DNA consisting of five genes) and a functional combined D-xylose utilization and zeaxanthin biosynthesis pathway (approximately 19 kb consisting of eight genes) with high efficiencies (70-100%) either on…

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Topics & keywords

Topics
Keywords
  • Biology
  • Functional genomics
  • DNA
  • Gene
  • Genetics
  • Saccharomyces cerevisiae
  • Plasmid
  • In vitro recombination
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