Rode C K, Obreque V H, Bloch C A
Department of Pediatrics, School of Medicine, University of Michigan, Ann Arbor 48109, USA.
Gene. 1995 Dec 1;166(1):1-9. doi: 10.1016/0378-1119(95)00630-5.
Genetic and biophysical techniques have traditionally been applied to genome mapping independently of one another. We present a series of Escherichia coli mini-Tn10 insertions that contain the rare-cutting polylinker 1 (RCP1) of rare restriction sites [including BlnI/AvrII, SpeI, NheI, XbaI, NotI, PacI and SfiI; Mahillon and Kleckner, Gene 116 (1992) 69-74] which allows them to be used not just for genetic mapping, but also for rapid physical mapping and integrated physical and genetic mapping of the E. coli chromosome. Their isolation and their physical and genetic coordinates in K-12 strain MG1655 are presented. Also, their use in purifying insertion-delimited DNAs from E. coli K-12 and in macrorestriction mapping of a pathogenic strain's chromosome is demonstrated. These insertions allow integration of (i) different macrorestriction patterns of a single strain's chromosome, (ii) the physical map of a single strain's chromosome with the genetic map of the species, and (iii) the physical maps of different strains' chromosomes.
传统上，基因技术和生物物理技术一直是彼此独立地应用于基因组作图。我们展示了一系列大肠杆菌mini-Tn10插入片段，它们含有稀有切割多克隆位点1（RCP1），该位点包含稀有限制位点[包括BlnI/AvrII、SpeI、NheI、XbaI、NotI、PacI和SfiI；Mahillon和Kleckner，《基因》116（1992）69 - 74]，这使得它们不仅可用于遗传作图，还可用于大肠杆菌染色体的快速物理作图以及物理和遗传综合作图。文中介绍了它们在K - 12菌株MG1655中的分离情况以及它们的物理和遗传坐标。此外，还展示了它们在从大肠杆菌K - 12中纯化插入片段界定的DNA以及在病原菌染色体的宏观限制酶切图谱绘制中的应用。这些插入片段能够实现以下整合：（i）单个菌株染色体的不同宏观限制酶切图谱；（ii）单个菌株染色体的物理图谱与物种遗传图谱；（iii）不同菌株染色体的物理图谱。
