Sanger F, Donelson J E, Coulson A R, Kössel H, Fischer D
Proc Natl Acad Sci U S A. 1973 Apr;70(4):1209-13. doi: 10.1073/pnas.70.4.1209.
A sequence of 50 residues in f1 DNA has been determined by the extension of a chemically synthesized octadeoxyribonucleotide by Escherichia coli DNA polymerase I, with radioactive nucleoside triphosphates and f1 DNA template. The polymerized product was synthesized either in the presence of manganese and a mixture of ribo- and deoxyribotriphosphates or in a magnesium-containing reaction with one or more of the four triphosphates absent. The sequence determination depended largely on fractionation of the polymerized products by two-dimensional "homochromatography." This approach and the techniques for the subsequent sequence analysis should be of general use for determining other sequences of DNA. Several features of this sequence suggest that it is located in an intercistronic region of f1 DNA.
利用放射性核苷三磷酸和f1 DNA模板，通过大肠杆菌DNA聚合酶I对化学合成的十八脱氧核糖核苷酸进行延伸，已确定了f1 DNA中一段50个残基的序列。聚合产物是在锰以及核糖和脱氧核糖三磷酸混合物存在的情况下合成的，或者是在缺少四种三磷酸之一或更多的含镁反应中合成的。序列测定主要依赖于通过二维“同系层析法”对聚合产物进行分级分离。这种方法以及后续序列分析的技术应该普遍适用于确定DNA的其他序列。该序列的几个特征表明它位于f1 DNA的基因间区域。
