THE PHYSICAL MAP OF THE WHOLE ESCHERICHIA-COLI CHROMOSOME - APPLICATION OF A NEW STRATEGY FOR RAPID ANALYSIS AND SORTING OF A LARGE GENOMIC LIBRARY
THE PHYSICAL MAP OF THE WHOLE ESCHERICHIA-COLI CHROMOSOME - APPLICATION OF A NEW STRATEGY FOR RAPID ANALYSIS AND SORTING OF A LARGE GENOMIC LIBRARY
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DOI：
10.1016/0092-8674(87)90503-4
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发表时间：
1987-07-31
期刊：
影响因子：
64.5
通讯作者：
ISONO, K
中科院分区：
文献类型：
--
作者：
KOHARA, Y;AKIYAMA, K;ISONO, K
3400个λ噬菌体克隆含有E.大肠杆菌染色体，并利用快速质谱分析法构建了一个4700 kb长的8个六碱基识别酶的整合限制性内切酶图谱。我们的策略是以类似于核苷酸测序的方式通过与载体探针杂交来测量部分限制性内切酶的大小。通过计算机程序将数据分组，并使用质量杂交方法进一步将边界克隆彼此关联。这些克隆可用于分离任何所需的E. coli基因，如果它们的图谱位置已知的话。此外，该策略也适用于其他生物的基因组分析。
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.