Singer M, Baker T A, Schnitzler G, Deischel S M, Goel M, Dove W, Jaacks K J, Grossman A D, Erickson J W, Gross C A
Microbiol Rev. 1989 Mar;53(1):1-24. doi: 10.1128/mr.53.1.1-24.1989.
We present a collection of 182 isogenic strains containing genetically linked antibiotic resistance elements located at approximately 1-min intervals around the Escherichia coli chromosome. At most positions both Tn10 (Tetr) and TN10kan (Kanr) elements are available, so that the collection contains a linked set of alternating antibiotic resistance markers. The map position of each insertion has been aligned to the E. coli genetic map as well as to the Kohara ordered clone bank. These strains are designed to be used in a rapid two-step mapping system in E. coli. In the first step, the mutation is localized to a 5- to 15-min region of the chromosome by Hfr mapping with a set of Hfr strains containing either Tn10 or Tn10kan elements located 20 min from their respective origins of transfer. In the second step, the mutation is localized to a 1-min region by P1 transduction, with a collection of isogenic insertion strains as donors. We discuss the uses of this collection of strains to map and eventually to clone a variety of mutations in E. coli.
我们展示了一组182个同基因菌株,这些菌株含有遗传连锁的抗生素抗性元件,它们围绕大肠杆菌染色体以大约1分钟的间隔分布。在大多数位置,Tn10(四环素抗性)和TN10kan(卡那霉素抗性)元件都可用,因此该菌株库包含一组连锁的交替抗生素抗性标记。每个插入位点的图谱位置已与大肠杆菌遗传图谱以及小原(Kohara)有序克隆文库进行了比对。这些菌株设计用于大肠杆菌的快速两步定位系统。第一步,通过用一组Hfr菌株进行Hfr定位,将突变定位到染色体的5至15分钟区域,这些Hfr菌株含有距离各自转移起点20分钟的Tn10或Tn10kan元件。第二步,通过P1转导,使用一组同基因插入菌株作为供体,将突变定位到1分钟区域。我们讨论了利用这组菌株对大肠杆菌中的各种突变进行定位并最终克隆的用途。