Effects of Infection with UV-or X-Ray-Irradiated T2 Phage Ghosts on RNA Synthesrs In Eschenchia coli

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Infection with UV-irradiated T2 phage ghosts stopped RNA synthesis in E. coli cells immediately and completely. X-ray-irradiated T2 phage lacked such function possibty due to decreased efflciency in adsorption. The effects of adsorption of T2 ghosts irradiated by UV light or X-rays on RNA synthesis of host E. coli cells were studied. It has been known that adsorption of T2 phage is not affected by UV Iight, but by X-rays. It has also been known that T2 phage ghosts stop immediately the RNA synthesis in host cells. A crude suspension of T2 phage was incubated with DNase and T2 phage were purified by differential centrifugation followed by centrifugation in a linear sucrosegradient (5-20%) in 4.4 ml of T2 buffer (3g Na_2HPO_4, 1.5 g KH_2PO_4, 4g NaCl, 5g K_2SO_4, 1 ml of 1 M MgSO4, 1 ml of 0.1M CaCl2 and 1 ml of 1% gelatin in 1,000 ml water) at 43,000xg for 25 min using a swinging bucket rotor. The fractions containing phage particles were collected and were dialysed against T2 buffer. An equal volume of glycerin was added to a dense suspension of T2 phage and mixed well. After 30 min incubation at room temperature, the suspension was diluted 40 fold with cold 1 mM Tris-buffer (pH 7. 4) with vigorous shaking. The mixture was then incubated for 30 min at 37℃ in the presence of 10 mM MgSO_4. The T2 ghosts were sedimented by a centrifugation at 23, 000xg for 1 h and were suspended in M9 medium (5.8g Na_2HPO_4.3g KH_2PO_4.1g NH_6Cl, 1 ml of 0,1 M CaCl_2.1 ml of 1 M MgSO_4, 10 ml of 40% glucose and 5 g NaCl in 1,000 ml mater) without glucose. The ghost-titer was determined by numbering E. coli cells at 37% survival fraction after mixing the ghosts with known numbers of bacterial cells.
日本放射線影響学会の論文

Effects of Infection with UV-or X-Ray-Irradiated T2 Phage Ghosts on RNA Synthesrs In Eschenchia coli

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