Umeda M, Ohtsubo E
Institute of Applied Microbiology, University of Tokyo, Japan.
J Mol Biol. 1989 Aug 20;208(4):601-14. doi: 10.1016/0022-2836(89)90151-4.
The chromosome of an Escherichia coli K-12 strain W3110 contains seven copies of insertion element IS1, 12 copies of IS2 and six copies of IS3. We determined the approximate locations of six copies of IS1 (named is1A to is1F), ten copies of IS2 (named is2A to is2J), and five copies of IS3 (named is3A to is3E) on the W3110 chromosome by plaque hybridization using the "mini-set" of the lambda phage library that includes 476 clones carrying chromosomal segments that cover the W3110 chromosome almost entirely. Cleavage maps of the W3110 chromosome and cleavage analysis of phage DNAs carrying insertion elements allowed us to assign more precise locations to most of the insertion elements and to determine their orientations. Insertion elements were distributed randomly along the W3110 chromosome in one or other orientation. Several of these were located at the same positions on the chromosome of another E. coli K-12 strain, JE5519, and they were assumed to be the original complement of insertion elements in E. coli K-12 wild-type. Locations and orientations of such insertion elements were correlated well with Hfr points of origin and with crossover points for excision of some F' factors derived from several Hfrs. Insertion elements may be involved also in rearrangement of bacterial chromosomes.
大肠杆菌K-12菌株W3110的染色体含有7个插入元件IS1拷贝、12个IS2拷贝和6个IS3拷贝。我们通过噬菌斑杂交,利用λ噬菌体文库的“微型集”确定了W3110染色体上6个IS1拷贝(命名为is1A至is1F)、10个IS2拷贝(命名为is2A至is2J)和5个IS3拷贝(命名为is3A至is3E)的大致位置,该文库包含476个携带染色体片段的克隆,这些片段几乎完全覆盖了W3110染色体。W3110染色体的切割图谱以及携带插入元件的噬菌体DNA的切割分析使我们能够为大多数插入元件确定更精确的位置并确定它们的方向。插入元件以一种或另一种方向沿着W3110染色体随机分布。其中有几个位于另一株大肠杆菌K-12菌株JE5519染色体的相同位置,它们被认为是大肠杆菌K-12野生型中插入元件的原始组成部分。这些插入元件的位置和方向与Hfr的起源点以及一些源自几个Hfr的F'因子切除的交叉点密切相关。插入元件也可能参与细菌染色体的重排。