Betlach M, Hershfield V, Chow L, Brown W, Goodman H, Boyer H W
Fed Proc. 1976 Jul;35(9):2037-43.
Genetic analyses of DNA restriction and modification mechanisms have been encumbered by the inability to rigorously select for mutant phenotypes associated with these systems. The application of restriction endonucleases has now proved to be a successful approach to the genetic analyses of small genomes that are recalcitrant to the more standard genetic techniques. Restriction endonucleases EcoRI and HindIII were used to analyze the structure of the plasmid genome responsible for the EcoRI restriction endonuclease and modification methylase. This plasmid in the original clinical isolate of Escherichia coli appears to be identical to the ColE 1 plasmid except for a 1.95 kilobase pair segment which contains these genes. A preliminary restriction map of this plasmid is presented.
DNA限制和修饰机制的遗传分析一直受到无法严格选择与这些系统相关的突变表型的阻碍。现在,限制内切酶的应用已被证明是对那些难以用更标准的遗传技术进行分析的小基因组进行遗传分析的成功方法。使用限制内切酶EcoRI和HindIII来分析负责EcoRI限制内切酶和修饰甲基化酶的质粒基因组的结构。除了包含这些基因的1.95千碱基对片段外,大肠杆菌原始临床分离株中的这种质粒似乎与ColE 1质粒相同。本文给出了该质粒的初步限制酶切图谱。