Phylogenetic screening of a bacterial, metagenomic library using homing endonuclease restriction and marker insertion.

ABSTRACT: Metagenomics provides access to the uncultured majority of the microbial world. The approaches employed in this field have, however, had limited success in linking functional genes to the taxonomic or phylogenetic origin of the organism they belong to. Here we present an efficient strategy to recover environmental DNA fragments that contain phylogenetic marker genes from metagenomic libraries. Our method involves the cleavage of 23S ribsosmal RNA (rRNA) genes within pooled library clones by the homing endonuclease I-CeuI followed by the insertion and selection of an antibiotic resistance cassette. This approach was applied to screen a library of 6500 fosmid clones derived from the microbial community associated with the sponge Cymbastela concentrica. Several fosmid clones were recovered af

SUBMITTER: Yung PY 

PROVIDER: S-EPMC2790883 | biostudies-literature | 2009 Nov

REPOSITORIES: biostudies-literature