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Characterization of Escherichia coli Single-gene Deletion Mutants Impaired in Bacteriophage ReproductionReimer, Kelly 27 June 2013 (has links)
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.
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Characterization of Escherichia coli Single-gene Deletion Mutants Impaired in Bacteriophage ReproductionReimer, Kelly 27 June 2013 (has links)
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.
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