title: Metagenomic analysis of Streptomyces lividans reveals host-dependent functional expression.
abstract: Most functional metagenomic studies have been limited by the poor expression of many genes derived from metagenomic DNA in Escherichia coli, which has been the predominant surrogate host to date. To expand the range of expressed genes, we developed tools for construction and functional screening of metagenomic libraries in Streptomyces lividans. We expanded on previously published protocols by constructing a system that enables retrieval and characterization of the metagenomic DNA from biologically active clones. To test the functionality of these methods, we constructed and screened two metagenomic libraries in S. lividans. One was constructed with pooled DNA from 14 bacterial isolates cultured from Alaskan soil and the second with DNA directly extracted from the same soil. Functional screening of these libraries identified numerous clones with hemolytic activity, one clone that produces melanin by a previously unknown mechanism, and one that induces the overproduction of a secondary metabolite native to S. lividans. All bioactive clones were functional in S. lividans but not in E. coli, demonstrating the advantages of screening metagenomic libraries in more than one host.
authors.affiliation: Department of Bacteriology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: McMahon
authors.forename: Changhui
authors.initials: C
authors.lastname: Guan
authors.forename: Jo
authors.initials: J
authors.lastname: Handelsman
authors.forename: Michael G
authors.initials: MG
authors.lastname: Thomas
mesh_terms: D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D020869:Gene Expression Profiling; D015723:Gene Library; D056186:Metagenomics / Q000379:methods*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D048170:Streptomyces lividans / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial