Sabik J F, Suit J L, Luria S E
J Bacteriol. 1983 Mar;153(3):1479-85. doi: 10.1128/jb.153.3.1479-1485.1983.
We isolated a series of Tn5 transposon insertion mutants and chemically induced mutants with mutations in the region of the ColE1 plasmid that includes the cea (colicin) and imm (immunity) genes. Bacterial cells harboring each of the mutant plasmids were tested for their response to the colicin-inducing agent mitomycin C. All insertion mutations within the cea gene failed to bring about cell killing after mitomycin C treatment. A cea- amber mutation exerted a polar effect on killing by mitomycin C. Two insertions beyond the cea gene but within or near the imm gene also prevented the lethal response to mitomycin C. These findings suggest the presence in the ColE1 plasmid of an operon containing the cea and kil genes whose product is needed for mitomycin C-induced lethality. Bacteria carrying ColE1 plasmids with Tn5 inserted within the cea gene produced serologically cross-reacting fragments of the colicin E1 molecule, the lengths of which were proportional to the distance between the insertion and the promoter end of the cea gene.
我们分离出了一系列Tn5转座子插入突变体以及在ColE1质粒中包含cea(大肠杆菌素)和imm(免疫)基因的区域发生突变的化学诱导突变体。对携带每种突变体质粒的细菌细胞进行了测试,以检测它们对大肠杆菌素诱导剂丝裂霉素C的反应。cea基因内的所有插入突变在丝裂霉素C处理后均未能导致细胞死亡。一个cea琥珀突变对丝裂霉素C诱导的杀伤作用产生了极性效应。在cea基因之外但在imm基因内或附近的两个插入也阻止了对丝裂霉素C的致死反应。这些发现表明ColE1质粒中存在一个含有cea和kil基因的操纵子,其产物是丝裂霉素C诱导致死性所必需的。携带在cea基因内插入Tn5的ColE1质粒的细菌产生了大肠杆菌素E1分子的血清学交叉反应片段,其长度与插入位点和cea基因启动子末端之间的距离成正比。