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Articles 1 - 3 of 3
Full-Text Articles in Biochemical and Biomolecular Engineering
Rapid Characterization And Engineering Of Natural Product Biosynthetic Pathways Via Dna Assembler, Zengyi Shao, Yunzi Luo, Huimin Zhao
Rapid Characterization And Engineering Of Natural Product Biosynthetic Pathways Via Dna Assembler, Zengyi Shao, Yunzi Luo, Huimin Zhao
Zengyi Shao
We report a synthetic biology strategy for rapid genetic manipulation of natural product biosynthetic pathways. Based on DNA assembler, this method synthesizes the entire expression vector containing the target biosynthetic pathway and the genetic elements required for DNA maintenance and replication in various hosts in a single-step manner through yeast homologous recombination, offering unprecedented flexibility and versatility in pathway manipulations.
Cloning And Heterologous Expression Of The Spectinabilin Biosynthetic Gene Cluster From Streptomyces Spectabilis, Zengyi Shao
Cloning And Heterologous Expression Of The Spectinabilin Biosynthetic Gene Cluster From Streptomyces Spectabilis, Zengyi Shao
Zengyi Shao
Spectinabilin is a rare nitrophenyl-substituted polyketide metabolite. Here we report the cloning and heterologous expression of the spectinabilin gene cluster from Streptomyces spectabilis. Unexpectedly, this gene cluster is evolutionarily closer to the aureothin gene cluster than to the spectinabilin gene cluster from Streptomyces orinoci. Moreover, the two nearly identical spectinabilin gene clusters use a distinctly different regulation mechanism.
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%) …