Biopharma Insights Here, we present phenomic approaches for the functional characterization of putative phage genes. Techniques include a developed assay capable of monitoring host anabolic metabolism, the Multi-phenotype Assay Plates (MAPs), in addition to the established method of metabolomics, capable of measuring effects to catabolic metabolism.
Total time
~5–7 days (includes plate preparation, continuous culture growth, sampling, and metabolomics analysis)
Model organism
Bacterial host (phage-susceptible strain, species not specified)
Steps
1
Prepare Multi-phenotype Assay Plates (MAPs)
Set up MAP assay plates with appropriate bacterial cultures and inoculation conditions to establish baseline anabolic metabolism measurements before phage exposure.
▶ 01:20
2
Measure Optical Density on MAP assays
Record optical density readings from MAP plates to quantify changes in host cell growth and anabolic metabolism in response to phage infection.
▶ 02:38
3
Set up chemostat continuous culture system
Initialize continuous culture bioreactors with sterile media, establish controlled growth conditions, and inoculate with bacterial cultures for metabolomics experiments.
▶ 03:27
4
Run continuous cultures for metabolomics sampling
Maintain steady-state continuous cultures, collect time-series samples following phage infection, and prepare samples for downstream metabolomics analysis of catabolic pathway changes.
▶ 05:07
5
Analyze phage gene phenotypes and metabolite profiles
Integrate MAP anabolic data and metabolomics results to functionally characterize putative phage genes and infer mechanisms of viral protein function.
▶ 07:15