DNA assembler: A synthetic biology tool for characterizing and engineering natural product gene clusters

Zengyi Shao, Huimin Zhao

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The majority of existing antibacterial and anticancer drugs are natural products or their derivatives. However, the characterization and engineering of these compounds are often hampered by limited ability to manipulate the corresponding biosynthetic pathways. Recently, we developed a genomics-driven, synthetic biology-based method, DNA assembler, for discovery, characterization, and engineering of natural product biosynthetic pathways (Shao, Luo, & Zhao, 2011). By taking advantage of the highly efficient yeast in vivo homologous recombination mechanism, this method synthesizes the entire expression vector containing the target biosynthetic pathway and the genetic elements needed for DNA maintenance and replication in individual hosts in a single-step manner. In this chapter, we describe the general guidelines for construct design. By using two distinct biosynthetic pathways, we demonstrate that DNA assembler can perform multiple tasks, including heterologous expression, introduction of single or multiple point mutations, scar-less gene deletion, generation of product derivatives, and creation of artificial gene clusters. As such, this method offers unprecedented flexibility and versatility in pathway manipulations.

Original languageEnglish (US)
Title of host publicationNatural Product Biosynthesis by Microorganisms and Plants, Part C
PublisherAcademic Press Inc.
Pages203-224
Number of pages22
ISBN (Print)9780124046344
DOIs
StatePublished - 2012

Publication series

NameMethods in Enzymology
Volume517
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Aureothin
  • Genetic manipulation of gene clusters
  • Heterologous expression
  • Natural product biosynthesis
  • Spectinabilin
  • Synthetic biology

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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  • Cite this

    Shao, Z., & Zhao, H. (2012). DNA assembler: A synthetic biology tool for characterizing and engineering natural product gene clusters. In Natural Product Biosynthesis by Microorganisms and Plants, Part C (pp. 203-224). (Methods in Enzymology; Vol. 517). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-404634-4.00010-3