Expression in Escherichia coli of chemiacally synthesized genes for human insulin

Goeddel, David V., et al. "Expression in Escherichia coli of chemically synthesized genes for human insulin." Proceedings of the National Academy of Sciences 76.1 (1979): 106-110. I actually had to read this paper for my lab report that I am doing this week so it does not have to do with RVLM, but still fun to know. This was actually one of the first studies to utilize protein recombination, and they did it in order to synthesize human insulin. At the time insulin was being harvested from animals at slow rates with some complication. Protein recombination allowed for cheap, fast, and large amounts of insulin production. In order to create the two insulin chains, a pIB1 plasmid and pIA1 plasmid were transformed into E. K-12 coli strain 294. After growth of the transformed bacteria, each colony was induced with IPTG for maximal expression of the insulin protein. The bacteria was then lysed and the mixture was S-sulfunated for the purification of the insulin chains. After purification a radioimmun assay and DNA sequencing was done for verification of the plasmid products. It was shown that human insulin had indeed been synthesized and was ready for wide spread use. ~JI