Publication Date

10-23-2024

Journal

Applied and Environmental Microbiology

DOI

10.1128/aem.01171-24

PMID

39315792

PMCID

PMC11497775

PubMedCentral® Posted Date

9-24-2024

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Gene Library, Bacteriophages, Metagenomics, Virome, Wastewater, Seawater, DNA, Viral, High-Throughput Nucleotide Sequencing, Genome, Viral, Fresh Water, Escherichia coli, metagenomic libraries, DNA libraries, virome, bacteriophage, barcoded DNA, pooled DNA libraries, metagenomics

Abstract

Despite many efforts to understand and leverage the functional potential of environmental viromes, most bacteriophage genes are largely uncharacterized. To explore novel biology from uncultivated microbes like phages, metagenomics has emerged as a powerful tool to directly mine new genes without the need to culture the diverse microbiota and the viruses within. When a pure computational approach cannot infer gene function, it may be necessary to create a DNA library from environmental genomic DNA, followed by the screening of that library for a particular function. However, these screens are often initiated without a metagenomic analysis of the completed DNA library being reported. Here, we describe the construction and characterization of DNA libraries from a single cultured phage (ΦT4), five cultured

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