Kohara Y, Akiyama K, Isono K
Cell. 1987 Jul 31;50(3):495-508. doi: 10.1016/0092-8674(87)90503-4.
Thirty-four hundred lambda phage clones containing segments of the E. coli chromosome were isolated and used to construct a 4700 kb long integrated restriction map for eight six-base-recognizing enzymes by a rapid mass-analysis method. Our strategy was to measure the sizes of partial restriction enzyme digests by hybridization with a vector probe in a manner analogous to nucleotide sequencing. The data were sorted into groups by a computer program and the boundary clones were further correlated with each other using a mass hybridization method. These clones can be exploited for the isolation of any desired E. coli genes if their map positions are known. Also, the strategy is applicable to analyses of the genomes of other organisms.
分离出3400个含有大肠杆菌染色体片段的λ噬菌体克隆,并通过一种快速批量分析方法,用于构建8种识别六碱基的酶的4700kb长的整合限制酶图谱。我们的策略是通过与载体探针杂交来测量部分限制酶消化产物的大小,其方式类似于核苷酸测序。数据通过计算机程序进行分组,边界克隆通过批量杂交方法进一步相互关联。如果知道这些克隆的图谱位置,就可以利用它们来分离任何所需的大肠杆菌基因。此外,该策略适用于其他生物基因组的分析。