Iowa State University From the SelectedWorks of Zengyi Shao 2009 DNA assembler , an in vivo genetic method for rapid construction of biochemical pathways
Iowa State University From the SelectedWorks of Zengyi Shao 2009 DNA assembler , an in vivo genetic method for rapid construction of biochemical pathways
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发表时间：
2016
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通讯作者：
Zengyi Shao;Hua Zhao;Huimin Zhao
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作者：
Zengyi Shao;Hua Zhao;Huimin Zhao
整个生化途径或基因组编码大型重组的DNA代表了一个重大挑战，我们报告了一种新方法，DNA组装器可以通过在体内重新组合sacCharomyces cerevisiae dna come dly dly comeyrers dly ryerer ryer ryer ryer ryer ryer ryer ryer ryer ryer ryer ryer save-new-new the the in in vivo。三个基因），一个功能性的Zeaxanthin生物合成途径（〜11 kb DNA由五个基因组成）和功能性的D- Xylose利用率和Zeaxanthin Biosynsisis途径（〜19 kB）（由八个基因组成），效率很高（70-100％）在质虫上的质量和元素均为ichomos siply and yeast and Oneptiation and Oneptiation。合成的生化途径的强大工具生物学，代谢工程和功能基因组学研究。
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.