Kadavy D R, Shaffer J J, Lott S E, Wolf T A, Bolton C E, Gallimore W H, Martin E L, Nickerson K W, Kokjohn T A
School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska 68588-0666, USA.
Appl Environ Microbiol. 2000 Dec;66(12):5206-12. doi: 10.1128/AEM.66.12.5206-5212.2000.
Reactivation of UV-C-inactivated Pseudomonas aeruginosa bacteriophages D3C3, F116, G101, and UNL-1 was quantified in host cells infected during the exponential phase, during the stationary phase, and after starvation (1 day, 1 and 5 weeks) under conditions designed to detect dark repair and photoreactivation. Our experiments revealed that while the photoreactivation capacity of stationary-phase or starved cells remained about the same as that of exponential-phase cells, in some cases their capacity to support dark repair of UV-inactivated bacteriophages increased over 10-fold. This enhanced reactivation capacity was correlated with the ca. 30-fold-greater UV-C resistance of P. aeruginosa host cells that were in the stationary phase or exposed to starvation conditions prior to irradiation. The dark repair capacity of P. aeruginosa cells that were infected while they were starved for prolonged periods depended on the bacteriophage examined. For bacteriophage D3C3 this dark repair capacity declined with prolonged starvation, while for bacteriophage G101 the dark repair capacity continued to increase when cells were starved for 24 h or 1 week prior to infection. For G101, the reactivation potentials were 16-, 18-, 10-, and 3-fold at starvation intervals of 1 day, 1 week, 5 weeks, and 1. 5 years, respectively. Exclusive use of exponential-phase cells to quantify bacteriophage reactivation should detect only a fraction of the true phage reactivation potential.
在旨在检测暗修复和光复活的条件下,对紫外线C灭活的铜绿假单胞菌噬菌体D3C3、F116、G101和UNL-1在指数生长期、稳定期以及饥饿(1天、1周和5周)后的宿主细胞中的复活情况进行了定量分析。我们的实验表明,虽然稳定期或饥饿细胞的光复活能力与指数生长期细胞的大致相同,但在某些情况下,它们支持紫外线灭活噬菌体暗修复的能力增加了10倍以上。这种增强的复活能力与处于稳定期或在照射前暴露于饥饿条件下的铜绿假单胞菌宿主细胞对紫外线C的抗性高约30倍有关。长时间饥饿时被感染的铜绿假单胞菌细胞的暗修复能力取决于所检测的噬菌体。对于噬菌体D3C3,这种暗修复能力随着饥饿时间的延长而下降,而对于噬菌体G101,当细胞在感染前饥饿24小时或1周时,暗修复能力继续增加。对于G101,在饥饿间隔1天、1周、5周和1.5年时的复活潜力分别为16倍、18倍、10倍和3倍。仅使用指数生长期细胞来定量噬菌体复活可能只能检测到真实噬菌体复活潜力的一部分。