Manipulating Natural Product Biosynthetic Pathways via DNA Assembler. Issue 2 (3rd June 2014)

• Record Type: Journal Article
• Authors: Shao, Zengyi; Zhao, Huimin
• Editors: Mahal, Lara; Romesberg, Floyd; Shah, Kavita; Shamu, Caroline; Strano, Michael S.; Thomas, Craig
• Is Part Of: Current protocols in chemical biology. Volume 6:Issue 2(2014)
• Page Start: 65
• Page End: 100
• Publication Date: 2014-06-03
• Subjects: natural product -- gene cluster -- regulation of natural product biosynthesis -- pathway engineering -- synthetic biology -- cryptic pathway -- DNA assembler; Biochemistry -- Laboratory manuals; Chemical Phenomena; Biochemistry; Periodicals; Laboratory manuals; 572.078
• DOI: 10.1002/9780470559277.ch130191
• Languages: English
• ISSNs: 2160-4762

Abstract

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. Curr. Protoc. Chem. Biol . 6:65‐100 © 2014 by John Wiley & Sons, Inc.