Phage phenomics [electronic resource] : Physiological approaches to characterize novel viral proteins

Published:: Washington, D.C. : United States. Dept. of Energy, 2015.
Oak Ridge, Tenn. : Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy
Physical Description:: Article numbers e52,854 : digital, PDF file
Additional Creators:: Argonne National Laboratory, United States. Department of Energy, and United States. Department of Energy. Office of Scientific and Technical Information

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Summary:

Current investigations into phage-host interactions are dependent on extrapolating knowledge from (meta)genomes. Interestingly, 60 - 95% of all phage sequences share no homology to current annotated proteins. As a result, a large proportion of phage genes are annotated as hypothetical. This reality heavily affects the annotation of both structural and auxiliary metabolic genes. Here we present phenomic methods designed to capture the physiological response(s) of a selected host during expression of one of these unknown phage genes. Multi-phenotype Assay Plates (MAPs) are used to monitor the diversity of host substrate utilization and subsequent biomass formation, while metabolomics provides bi-product analysis by monitoring metabolite abundance and diversity. Both tools are used simultaneously to provide a phenotypic profile associated with expression of a single putative phage open reading frame (ORF). Thus, representative results for both methods are compared, highlighting the phenotypic profile differences of a host carrying either putative structural or metabolic phage genes. In addition, the visualization techniques and high throughput computational pipelines that facilitated experimental analysis are presented.

Report Numbers:

E 1.99:1224998

Subject(s):

Basic Biological Sciences

Note:

Published through SciTech Connect.
06/11/2015.
Journal of Visualized Experiments 100 ISSN 1940-087X AM
Sanchez, Savannah; Cuevas, Daniel; Rostron, Jason; Liang, Tiffany; Pivaroff, Cullen; Haynes, Matthew; Nulton, Jim; Felts, Ben; Bailey, Barbara; Salamon, Peter; Edwards, Robert; Burgin, Alex; Segall, Anca; Rohwer, Forest.

Funding Information:

AC02-06CH11357

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