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紫外线照射的T4噬菌体DNA的部分复制导致特定基因区域的扩增。

Partial replication of UV-irradiated T4 bacteriophage DNA results in amplification of specific genetic areas.

作者信息

Ling S K, Vogelbacker H H, Restifo L L, Mattson T, Kozinski A W

出版信息

J Virol. 1981 Nov;40(2):403-10. doi: 10.1128/JVI.40.2.403-410.1981.

Abstract

Upon infection of Escherichia coli with bromodeoxyuridine-labeled t4 phage that had received 10 lethal hits of UV irradiation, a sizable amount of phage DNA was synthesized (approximately 36 phage equivalent units of DNA per infected bacterium), although very little multiplicity reactivation occurs. This progeny DNA was isolated and analyzed. This DNA was biased in its genetic representation, as shown by hybridization to cloned segments of the T4 genome immobilized on nitrocellulose filters. Preferentially amplified areas corresponded to regions containing origins of T4 DNA replication. The size of the progeny DNA increased with time after infection, possibly due to recombination between partial replicas and nonreplicated subunits or due to the gradual overcoming of the UV damage. As the size of the progeny DNA increased, all of the genes were more equally represented, resulting in a decrease in the genetic bias. Amplification of specific genetic areas was also observed upon infection with UV-irradiated, nonbromodeoxyuridine-substituted (light) phage. However, the genetic bias observed in this case was not as great as that observed with bromodeoxyuridine-substituted phage. This is most likely due to the higher efficiency of multiplicity reactivation of the light phage.

摘要

用经紫外线照射遭受10次致死剂量打击且用溴脱氧尿苷标记的T4噬菌体感染大肠杆菌后,尽管多重复活极少发生,但仍合成了大量噬菌体DNA(每个受感染细菌约有36个噬菌体等效单位的DNA)。分离并分析了这种子代DNA。如与固定在硝酸纤维素滤膜上的T4基因组克隆片段杂交所示,这种DNA在基因代表性上存在偏差。优先扩增的区域对应于含有T4 DNA复制起点的区域。子代DNA的大小在感染后随时间增加,这可能是由于部分复制体与未复制亚基之间的重组,或者是由于紫外线损伤的逐渐克服。随着子代DNA大小的增加,所有基因的代表性更加平均,导致基因偏差减小。在用紫外线照射、未用溴脱氧尿苷取代的(轻)噬菌体感染时,也观察到特定基因区域的扩增。然而,在这种情况下观察到的基因偏差不如用溴脱氧尿苷取代的噬菌体那么大。这很可能是由于轻噬菌体多重复活的效率更高。

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