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克隆的噬菌体φX174基因E的裂解作用

Lytic action of cloned phi X174 gene E.

作者信息

Young K D, Young R

出版信息

J Virol. 1982 Dec;44(3):993-1002. doi: 10.1128/JVI.44.3.993-1002.1982.

Abstract

The phi X174 lysis gene E was placed under control of the lac promoter by cloning into the multicopy plasmid pBH20. Other phi X174 gene sequences were removed by nuclease digestion. Expression of gene E was shown to be necessary and sufficient to produce lysis phenomena exhibited by infection with intact phage. Lysis, its inhibition by MgSO4 and spermine, its progression through a spheroplasting stage, and its dependence on an early chloramphenicol-sensitive step were reproduced in clones induced for expression of the E gene product. Escherichia coli clones carrying the E gene not under lac control, and clones under lac control but only minimally induced for gene E expression, exhibited morphological aberrations consistent with the view that the mechanism by which gene E mediates cell lysis is related to host cell division processes.

摘要

通过克隆到多拷贝质粒pBH20中,将噬菌体φX174的裂解基因E置于乳糖启动子的控制之下。通过核酸酶消化去除了其他φX174基因序列。结果表明,基因E的表达对于产生完整噬菌体感染所表现出的裂解现象是必要且充分的。在诱导表达E基因产物的克隆中,重现了裂解现象、其被硫酸镁和精胺抑制的现象、其通过原生质球化阶段的进程以及其对早期氯霉素敏感步骤的依赖性。携带不受乳糖控制的E基因的大肠杆菌克隆,以及受乳糖控制但仅对基因E表达进行最小诱导的克隆,表现出形态异常,这与基因E介导细胞裂解的机制与宿主细胞分裂过程相关的观点一致。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5c33/256359/4fb78efd411c/jvirol00153-0241-a.jpg

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