An assay was designed to measure the sensitivity of Escherichia coli mutants to bacteriophage infection via growth curves, using a Tecan temperature controlled plate reader. I screened 3985 single-gene deletion strains in Escherichia coli K12 from the Keio collection and identified 43 strains displaying varying degrees of resistance to four different phages, three non-contractile tailed phages (λcI857, HK97, and HK243) and the myoviridae T6, including 20 genes not previously implicated in phage infection. Additional assays, such as adsorption and tests of DNA-injection, were designed to further characterize resistant strains. The use of these assays helped identify varying sensitivities to LPS structure and LamB receptor concentration in the three non-contractile tailed phages, showing HK97 is the most sensitive to changes and HK243 the least. I also found that the periplasmic chaperone, FkpA, is required for HK97 DNA-injection.
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/35522 |
| Date | 27 June 2013 |
| Creators | Reimer, Kelly |
| Contributors | Davidson, Alan R., Edwards, Aled M. |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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