Moore S K, James E
Gene. 1979 Feb;5(2):159-75. doi: 10.1016/0378-1119(79)90100-8.
A detailed physical map depicting the cleavage sites generated by ten different restriction endonucleases was prepared for the argF region of the Escherichia coli K-12 genome carried on a 1650 base pair fragment capable of directing the in vitro synthesis of ornithine transcarbamylase (OTCase; ec 2.1.3.3) under the control of arginine holorepressor. The method employed was originally developed by Smith and Birnstiel (1976), and involved the electrophoretic sizing of partial endonuclease digestion products of DNA radiolabeled at one end. This novel technique proved to be rapid, simple, amenable to the simultaneous mapping of numerous cleavage sites, and provided the essential information for determining the map order of restriction fragments. A facile method which involved magnesium phosphate as the DNA-binding agent was presented for the isolation of DNA fragments. The discovery of a 117 base pair leader sequence in the argF gene is also discussed.
针对大肠杆菌K-12基因组的argF区域，制备了一幅详细的物理图谱，该图谱描绘了由十种不同限制性内切酶产生的切割位点。该argF区域位于一个1650碱基对的片段上，此片段能够在精氨酸全阻遏物的控制下指导鸟氨酸转氨甲酰酶（OTCase；ec 2.1.3.3）的体外合成。所采用的方法最初由史密斯和伯恩斯蒂尔于1976年开发，包括对一端进行放射性标记的DNA的部分内切酶消化产物进行电泳大小测定。这种新技术被证明是快速、简单的，适用于同时绘制多个切割位点的图谱，并为确定限制性片段的图谱顺序提供了重要信息。还提出了一种以磷酸镁作为DNA结合剂的简便方法来分离DNA片段。文中还讨论了在argF基因中发现的一个117碱基对的前导序列。
