Manipulating natural product biosynthetic pathways via DNA assembler

Zengyi Shao, Huimin Zhao

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)65-100
Number of pages36
JournalCurrent protocols in chemical biology
Volume6
Issue number2
DOIs
StatePublished - 2014

Keywords

  • DNA assembler
  • cryptic pathway
  • gene cluster
  • natural product
  • pathway engineering
  • regulation of natural product biosynthesis
  • synthetic biology

ASJC Scopus subject areas

  • Medicine(all)

Fingerprint Dive into the research topics of 'Manipulating natural product biosynthetic pathways via DNA assembler'. Together they form a unique fingerprint.

Cite this