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Full-Text Articles in Engineering
Activation And Characterization Of A Cryptic Polycyclic Tetramate Macrolactam Biosynthetic Gene Cluster, Yunzi Luo, Hua Huang, Jing Liang, Meng Wang, Lu Lu, Zengyi Shao, Ryan E. Cobb, Huimin Zhao
Activation And Characterization Of A Cryptic Polycyclic Tetramate Macrolactam Biosynthetic Gene Cluster, Yunzi Luo, Hua Huang, Jing Liang, Meng Wang, Lu Lu, Zengyi Shao, Ryan E. Cobb, Huimin Zhao
Zengyi Shao
Polycyclic tetramate macrolactams (PTMs) are a widely distributed class of natural products with important biological activities. However, many of these PTMs have not been characterized. Here we apply a plug-and-play synthetic biology strategy to activate a cryptic PTM biosynthetic gene cluster SGR810-815 from Streptomyces griseus and discover three new PTMs. This gene cluster is highly conserved in phylogenetically diverse bacterial strains and contains an unusual hybrid polyketide synthase-nonribosomal peptide synthetase, which resembles iterative polyketide synthases known in fungi. To further characterize this gene cluster, we use the same synthetic biology approach to create a series of gene deletion constructs and …
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.
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%) …