Birkenbihl R P, Vielmetter W
Institut für Genetik, Universität zu Köln, FRG.
Nucleic Acids Res. 1989 Jul 11;17(13):5057-69. doi: 10.1093/nar/17.13.5057.
A physical map for the genome of E. coli K12 strain BHB2600 was constructed by use of 570 cloned DNA elements (CDEs) withdrawn from a cosmid library. Dot blot hybridisation was applied to establish contig interrelations with subsequent fine mapping achieved by analysis of EcoR1 restriction patterns on Southern blots. The derived map covers nearly 95% of the E. coli genome resulting in 12 minor gaps. It may be compared to the almost complete map for strain W3110 of Kohara et al. (1). Except for one tiny gap (lpp,36.5') remaining gaps in BHB2600 do not coincide with those in W3110 so that both maps complement each other establishing an essentially complete clone represented map. Besides numerous minute differences (site and fragment gains and losses) both strains harbour at differing positions extended rearrangements flanked by mutually inverted repetitive elements, in our case insertion elements (IS1 and IS5).
利用从黏粒文库中提取的570个克隆DNA元件(CDEs)构建了大肠杆菌K12菌株BHB2600的物理图谱。通过斑点印迹杂交来建立重叠群的相互关系,随后通过分析Southern印迹上的EcoR1限制酶切图谱实现精细定位。得到的图谱覆盖了近95%的大肠杆菌基因组,产生了12个小缺口。它可以与Kohara等人(1)构建的W3110菌株的几乎完整的图谱相比较。除了一个微小缺口(lpp,36.5')外,BHB2600中其余的缺口与W3110中的缺口不一致,因此这两个图谱相互补充,形成了一个基本完整的克隆代表图谱。除了许多微小差异(位点和片段的得失)外,这两个菌株在不同位置都存在由相互反向的重复元件(在我们的例子中是插入元件(IS1和IS5))侧翼的延伸重排。