Xiong G, Oepen P, Geiben R, el-Idrissi A H, Lutz F
Institut für Pharmakologie und Toxikologie, Justus-Liebig-Universität Giessen, Germany.
Virus Res. 1996 Mar;41(1):77-87. doi: 10.1016/0168-1702(95)01279-6.
In bacteriophage phi CTX, the cohesive end sequences cos, the integrase gene int, the attachment site attP (the target site for int) and the gene ctx encoding a pore-forming cytotoxin CTX, are clustered. Phi CTX can infect some Pseudomonas aeruginosa strains with a subsequent induction of CTX expression. The 41 and 477 bp fragments containing cos ends of phi CTX DNA were cloned into the high copy number plasmid pHA10. After pretransformation with the cos ends containing plasmids, plaque formation of phi CTX and cytotoxin production in phi CTX-infected Pseudomonas aeruginosa cells decreased by 100- and 10-fold respectively. Twelve hours after phi CTX infection proteins binding with cooperativity to cos sequence containing cleavable ends and the 10 bp flanking sequences were detected by gel electrophoretic mobility retardation of [32P]cos DNA. The results suggest that the cos binding proteins of phi CTX are involved in phi CTX replication.
在噬菌体phi CTX中,粘性末端序列cos、整合酶基因int、附着位点attP(int的靶位点)以及编码成孔细胞毒素CTX的基因ctx聚集在一起。Phi CTX可以感染一些铜绿假单胞菌菌株,并随后诱导CTX表达。将含有phi CTX DNA cos末端的41和477 bp片段克隆到高拷贝数质粒pHA10中。在用含有cos末端的质粒进行预转化后,phi CTX的噬菌斑形成以及phi CTX感染的铜绿假单胞菌细胞中的细胞毒素产生分别降低了100倍和10倍。在phi CTX感染12小时后,通过[32P]cos DNA的凝胶电泳迁移率阻滞检测到与含有可切割末端的cos序列和10 bp侧翼序列协同结合的蛋白质。结果表明,phi CTX的cos结合蛋白参与了phi CTX的复制。