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PMID: 24903884 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Manipulating natural product biosynthetic pathways via DNA assembler.

Current protocols in chemical biology ·Vol. 6 ·No. 2 ·2014-06-03 ·Pages 65-100

Shao Z, Zhao H

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.

Keywords
DNA assembler cryptic pathway gene cluster natural product pathway engineering regulation of natural product biosynthesis synthetic biology
MeSH Terms
Antimalarials Antiviral Agents Biological Products/chemistry Biosynthetic Pathways/genetics Cloning, Molecular DNA/chemistry DNA Primers Deoxyribonucleases/chemistry Gene Deletion Multigene Family Pyrones Streptomyces/chemistry,genetics Streptomyces lividans/genetics,metabolism
Chemicals
Antimalarials Antiviral Agents Biological Products DNA Primers Pyrones spectinabilin DNA Deoxyribonucleases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Shao Zengyi
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.
Zhao Huimin
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.
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Article Info
Journal
Current protocols in chemical biology
Abbr.
Curr Protoc Chem Biol
ISSN
2160-4762
DOI
10.1002/9780470559277.ch130191
Published
2014-06-03
Epub
2014-00-03
Pages
65-100
Language
English
Region
United States
NLM ID
101559235
PMCID
PMC4105043
Subset
IM
Grants
NIGMS NIH HHS · P01 GM077596 · United States
NIGMS NIH HHS · GM077596 · United States
External Links
PubMed source DOI: 10.1002/9780470559277.ch130191 PMC Full Text
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