Genome-Wide High-Throughput Mining of Natural-Product Biosynthetic Gene Clusters by Phage Display

Jun Yin, Paul D. Straight, Siniša Hrvatin, Pieter C. Dorrestein, Stefanie B. Bumpus, Cindy Jao, Neil L Kelleher, Roberto Kolter*, Christopher T. Walsh

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

46 Scopus citations

Abstract

We have developed a phage-display method for high-throughput mining of bacterial gene clusters encoding the natural-product biosynthetic enzymes, polyketide synthases (PKSs) and nonribosomal peptide synthetases (NRPSs). This method uses the phosphopantetheinyl transferase activity of Sfp to specifically biotinylate NRPS and PKS carrier-protein domains expressed from a library of random genome fragments fused to a gene encoding a phage coat protein. Subsequently, the biotinylated phages are enriched through selection on streptavidin-coated plates. Using this method, we isolated phage clones from the multiple NRPS and PKS gene clusters encoded in the genomes of Bacillus subtilis and Myxococcus xanthus. Due to the rapid and unambiguous identification of carrier domains, this method will provide an efficient tool for high-throughput cloning of NRPS and PKS gene clusters from many individual bacterial genomes and multigenome environmental DNA.

Original languageEnglish (US)
Pages (from-to)303-312
Number of pages10
JournalChemistry and Biology
Volume14
Issue number3
DOIs
StatePublished - Mar 2007

Keywords

  • CHEMBIOL
  • MICROBIO

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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