Selective Enrichment of Environmental DNA Libraries for Genes Encoding Nonribosomal Peptides and Polyketides by Phosphopantetheine Transferase-Dependent Complementation of Siderophore Biosynthesis

被引：16

作者：

Charlop-Powers, Zachary ^{[1
]}

Banik, Jacob J. ^{[1
]}

Owen, Jeremy G. ^{[1
]}

Craig, Jeffrey W. ^{[1
]}

Brady, Sean F. ^{[1
]}

机构：

[1] Rockefeller Univ, Howard Hughes Med Inst, Lab Genetically Encoded Small Mol, New York, NY 10065 USA

来源：

ACS CHEMICAL BIOLOGY | 2013年 / 8卷 / 01期

基金：

美国国家卫生研究院;

关键词：

NATURAL-PRODUCTS; GENOME; IDENTIFICATION; ENTEROBACTIN; SUPERFAMILY; DIVERSITY; CLUSTERS;

D O I：

10.1021/cb3004918

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The cloning of DNA directly from environmental samples provides a means to functionally access biosynthetic gene clusters present in the genomes of the large fraction of bacteria that remains recalcitrant to growth in the laboratory. Herein, we demonstrate a method by which complementation of phosphopantetheine transferase deletion mutants can be used to restore siderophore biosynthesis and to therefore selectively enrich eDNA libraries for nonribosomal peptide synthetase (NAPS) and polyketide synthase (PKS) gene sequences to unprecedented levels. The common use of NRPS/PKS-derived siderophores across bacterial taxa makes this method generalizable and should allow for the facile selective enrichment of NRPS/PKS-containing biosynthetic gene clusters from large environmental DNA libraries using a wide variety of phylogenetically diverse bacterial hosts.

引用

页码：138 / 143

页数：6