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Monitoring Infection of E. Coli By Bacteriophage T7 through Mass Spectrometry-Based Proteomics
Schienschang, Amy Swayne
Schienschang, Amy Swayne
Abstract
Mass spectrometry-based proteomics has become an important and versatile tool in analytical chemistry, making sense of complex biological samples and shedding light on the intricate proteomes of living organisms. Bottom-up proteomics studies are used to elucidate the changes in gene expression of bacteriophage T7 over the course of infection of Escherichia coli. E. coli cultures were infected with T7, sampled over time, and proteins were isolated and enzymatically digested. Nanoflow liquid chromatography combined with tandem mass spectrometry was used to detect proteolytic peptides and identify host and phage proteins. Generally, phage proteins were detected on a time scale fitting the established lytic cycle for T7 phage, confirming the effectiveness of infection monitoring by mass spectrometry-based proteomics studies. Continued development of the experimental method sought to increase detection of proteolytic peptides and identify phage and host proteins to a higher level of confidence, and lead to the implementation of 1D SDS-PAGE as a fractionation method to reduce sample complexity and increase method sensitivity.
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2018-01-01
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Chemistry
DOI
http://dx.doi.org/10.21220/s2-jfyf-3r27