Genome-wide high-throughput mining of natural-product biosynthetic gene clusters by phage display. Academic Article

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

• Straight, Paul

published proceedings

• Chem Biol

altmetric score

• 10

author list (cited authors)

• Yin, J., Straight, P. D., Hrvatin, S., Dorrestein, P. C., Bumpus, S. B., Jao, C., ... Walsh, C. T.

citation count

• 52

complete list of authors

• Yin, Jun||Straight, Paul D||Hrvatin, Sinisa||Dorrestein, Pieter C||Bumpus, Stefanie B||Jao, Cindy||Kelleher, Neil L||Kolter, Roberto||Walsh, Christopher T

publication date

• January 2007

publisher

• Elsevier  Publisher

published in

• Chemistry and Biology  Journal

keywords

• Bacillus Subtilis
• Bacterial Proteins
• Genes, Synthetic
• Multigene Family
• Myxococcus Xanthus
• Peptide Library
• Peptide Synthases
• Polyketide Synthases

Digital Object Identifier (DOI)

• 10.1016/j.chembiol.2007.01.006

start page

• 303

end page

• 312

volume

• 14

issue

• 3

URL

• http://dx.doi.org/10.1016/j.chembiol.2007.01.006