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Dna Assembler, An In Vivo Genetic Method For Rapid Construction Of Biochemical Pathways, Zengyi Shao, Hua Zhao, Huimin Zhao
Dna Assembler, An In Vivo Genetic Method For Rapid Construction Of Biochemical Pathways, Zengyi Shao, Hua Zhao, Huimin Zhao
Zengyi Shao
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%) …