Bläsi U, Henrich B, Lubitz W
J Gen Microbiol. 1985 May;131(5):1107-14. doi: 10.1099/00221287-131-5-1107.
The lysis gene E of bacteriophage phi X174 was cloned under transcriptional control of the lefthanded lambda promoter, giving rise to plasmid pSB12. Plasmid pSB22, identical to pSB12 except for an amber mutation in gene E, was constructed in the same way. Induction of the cloned wild-type gene by heat inactivation of the thermosensitive lambda cI857 repressor resulted in lysis of the host bacteria. With plasmid pSB22 only amber suppressor strains of Escherichia coli lysed after heat inactivation of lambda cI857. Lysis of E. coli was shown to depend on the rate of gene E translation and on the growth phase of the bacteria. Stationary cells could not be lysed by the gene E product (gpE), even if present in sufficient amounts to lyse growing cells. By isotopic labelling gpE could be detected among the proteins synthesized in normal E. coli as well as in minicells. Determination of gene E expression suggested that gpE synthesis is translationally regulated.
噬菌体φX174的裂解基因E在左手λ启动子的转录控制下进行克隆,从而产生了质粒pSB12。质粒pSB22的构建方式与pSB12相同,只是基因E中有一个琥珀突变。通过热敏感λcI857阻遏物的热失活来诱导克隆的野生型基因,会导致宿主细菌裂解。对于质粒pSB22,只有在λcI857热失活后,大肠杆菌的琥珀抑制菌株才会裂解。已证明大肠杆菌的裂解取决于基因E的翻译速率和细菌的生长阶段。即使基因E产物(gpE)的量足以裂解生长中的细胞,静止细胞也不会被其裂解。通过同位素标记,在正常大肠杆菌以及微细胞中合成的蛋白质中都能检测到gpE。对基因E表达的测定表明,gpE的合成受到翻译调控。