Johnson D A, Willetts N S
J Bacteriol. 1980 Sep;143(3):1171-8. doi: 10.1128/jb.143.3.1171-1178.1980.
The largest R . BamHI fragment of the plasmid F, which carries the entire F conjugation system, has been cloned into the single R . BamHI site of the ampicillin (Ap) resistance transposon TN1. pDS1106 (ColE1 mob::Tn1) was the vector plasmid, and the resultant conjugative plasmid, pED830, was characterized both genetically and by restriction enzyme analysis. The transposon construct, denoted Tn2301, was transposable at frequencies similar to Tn1 to small nonconjugative plasmids or to the Escherichia coli host chromosome. In the former case, Apr conjugative plasmids were obtained, whereas in the latter case, Hfr strains resulted. Representative Hfr strains were characterized by quantitative and interrupted mating experiments. Extension of this technique for Hfvr formation should aid chromosome mapping both in E. coli and in other bacterial genera.
携带完整F接合系统的质粒F的最大R. BamHI片段已被克隆到氨苄青霉素（Ap）抗性转座子TN1的单个R. BamHI位点中。pDS1106（ColE1 mob::Tn1）是载体质粒，通过遗传分析和限制性酶切分析对所得的接合性质粒pED830进行了表征。所构建的转座子，命名为Tn2301，其转座频率与Tn1相似，可转移到小型非接合性质粒或大肠杆菌宿主染色体上。在前一种情况下，可获得Apr接合性质粒，而在后一种情况下，则产生高频重组（Hfr）菌株。通过定量和中断杂交实验对代表性的Hfr菌株进行了表征。这种用于形成Hfr的技术的扩展应该有助于大肠杆菌和其他细菌属的染色体图谱绘制。
