title: Genome-wide high-throughput mining of natural-product biosynthetic gene clusters by phage display.
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.
authors: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
authors: Jun
authors: J
authors: Yin
authors: Paul D
authors: PD
authors: Straight
authors: Sinisa
authors: S
authors: Hrvatin
authors: Pieter C
authors: PC
authors: Dorrestein
authors: Stefanie B
authors: SB
authors: Bumpus
authors: Cindy
authors: C
authors: Jao
authors: Neil L
authors: NL
authors: Kelleher
authors: Roberto
authors: R
authors: Kolter
authors: Christopher T
authors: CT
authors: Walsh
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D005813:Genes, Synthetic; D005810:Multigene Family; D016941:Myxococcus xanthus / Q000201:enzymology / Q000235:genetics; D019151:Peptide Library; D010453:Peptide Synthases / Q000235:genetics; D048630:Polyketide Synthases / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D019151:Peptide Library; D048630:Polyketide Synthases; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase