title: Manipulating natural product biosynthetic pathways via DNA assembler.
abstract: DNA assembler is an efficient synthetic biology method for constructing and manipulating biochemical pathways. The rapidly increasing number of sequenced genomes provides a rich source for discovery of gene clusters involved in synthesizing new natural products. However, both discovery and economical production are hampered by our limited knowledge in manipulating most organisms and the corresponding pathways. By taking advantage of yeast in vivo homologous recombination, DNA assembler synthesizes an entire expression vector containing the target biosynthetic pathway and the genetic elements needed for DNA maintenance and replication. Here we use the spectinabilin clusters originated from two hosts as examples to illustrate the guidelines of using DNA assembler for cluster characterization and silent cluster activation. Such strategies offer unprecedented versatility in cluster manipulation, bypass the traditional laborious strategies to elicit pathway expression, and provide a new platform for de novo cluster assembly and genome mining for discovering new natural products.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois.|Department of Chemical and Biological Engineering, Iowa State University, Ames, Iowa.
authors.forename: Zengyi
authors.initials: Z
authors.lastname: Shao
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois.|Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois.|Departments of Chemistry, Biochemistry, and Bioengineering, University of Illinois at Urbana-Champaign, Urbana, Illinois.
authors.forename: Huimin
authors.initials: H
authors.lastname: Zhao
mesh_terms: D000962:Antimalarials; D000998:Antiviral Agents; D001688:Biological Products / Q000737:chemistry*; D053898:Biosynthetic Pathways / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry*; D017931:DNA Primers; D003851:Deoxyribonucleases / Q000737:chemistry; D017353:Gene Deletion; D005810:Multigene Family; D011753:Pyrones; D013302:Streptomyces / Q000737:chemistry / Q000235:genetics; D048170:Streptomyces lividans / Q000235:genetics / Q000378:metabolism
chemical_list: D000962:Antimalarials; D000998:Antiviral Agents; D001688:Biological Products; D017931:DNA Primers; D011753:Pyrones; C508401:spectinabilin; D004247:DNA; D003851:Deoxyribonucleases
keywords: DNA assembler; cryptic pathway; gene cluster; natural product; pathway engineering; regulation of natural product biosynthesis; synthetic biology