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Sanger, a physician's son from Rendcombe in England, received both his BA and his PhD from Cambridge University (in 1939 and 1943 respectively). He continued his research at the university and from 1951 until 1983 was a member of the scientific staff of the Medical Research Council. In 1955, after some ten years' work, Sanger established the complete amino-acid sequence of the protein bovine insulin. This was one of the first protein structures identified, and Sanger received the Nobel Prize for chemistry in 1958 in recognition of his achievement. Sanger's work enabled chemists to synthesize insulin artificially and generally stimulated research in protein structure.

In 1977 Sanger's team at the MRC laboratories, Cambridge, published the complete nucleotide (base) sequence of the genetic material (DNA) of the virus Phi X 174. This involves determining the order of 5400 nucleotides along the single circular DNA strand. Moreover they found two cases of genes located within genes. Previously it had been thought that genes could not overlap. Sanger's research required the development of new techniques for splitting the DNA into different-sized fragments. These are radioactively labeled and then separated by electrophoresis. The base sequence can then be worked out because it is known which base is located at the end of each fragment due to the specificity of the enzymes (the so-called restriction enzymes) used to split the DNA. Sanger was awarded the Nobel Prize for chemistry a second time (1980) for his work on determining the base sequences of nucleic acids.