Novel 'Bacteriospray' Method Facilitates the Functional Screening of Metagenomic Libraries for Antimicrobial Activity.

ABSTRACT: Metagenomic techniques, notably the cloning of environmental DNA (eDNA) into surrogate hosts, have given access to the genome of uncultured bacteria. However, the determination of gene functions based on DNA sequences alone remains a significant challenge. The functional screening of metagenomic libraries represents an interesting approach in the discovery of microbial metabolites. We describe here an optimized screening approach that facilitates the identification of new antimicrobials among large metagenomic libraries. Notably, we report a detailed genomic library construction protocol using Escherichia coli DH10B as a surrogate host, and demonstrate how vector/genomic DNA dephosphorylation, ligase inactivation, dialysis of the ligation product and vector/genomic DNA ratio greatly influe

SUBMITTER: Brahami A 

PROVIDER: S-EPMC6481063 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature