Russell Grimwade School of Biochemistry, University of Melbourne, Parkville, Victoria 3052, and Division of Food Processing, Dairy Research Laboratory, Commonwealth Scientific and Industrial Research Organization, P.O. Box 20, Highett, Victoria 3190, Australia.
Appl Environ Microbiol. 1988 Apr;54(4):1039-45. doi: 10.1128/aem.54.4.1039-1045.1988.
The temperate bacteriophage BK5-T was isolated from Streptococcus cremoris BK5 by induction with mitomycin C. Electron microscopy revealed that BK5-T DNA consists of linear molecules, ranging in size from 39.7 to 46 kilobase pairs. Restriction analysis of self-ligated BK5-T DNA showed that the ends of the DNA were not cohesive. The EcoRI restriction fragments of the phage genome were cloned into pACYC184. Restriction enzyme analysis of both the phage DNA and the cloned EcoRI fragments with EcoRI, BstEII, PstI, ClaI, and XbaI yielded a 37.6-kilobase-pair-long circular restriction map for the phage genome. It was concluded that the BK5-T DNA molecules in the population differ in their sequence by a circular permutation and that individual DNA molecules are terminally redundant. The map location of the sites at which packaging of BK5-T DNA into phage heads is initiated (pac) and at which the phage integrates into the bacterial chromosome (att) were established.
温和噬菌体 BK5-T 是通过丝裂霉素 C 诱导从乳球菌 BK5 中分离出来的。电子显微镜显示,BK5-T DNA 由线性分子组成,大小从 39.7 到 46 千碱基对不等。自连接 BK5-T DNA 的限制分析表明,DNA 的末端不具有粘性。噬菌体基因组的 EcoRI 限制片段被克隆到 pACYC184 中。用 EcoRI、BstEII、PstI、ClaI 和 XbaI 对噬菌体 DNA 和克隆的 EcoRI 片段进行酶切分析,得到了噬菌体基因组的 37.6 千碱基对长的圆形限制图谱。结论是,群体中的 BK5-T DNA 分子在序列上存在环状排列差异,并且单个 DNA 分子在末端是冗余的。确定了包装 BK5-T DNA 进入噬菌体头部的起始位点 (pac) 和噬菌体整合到细菌染色体上的位点 (att) 的位置。