Expression in Escherichia coli of chemically synthesized genes for human insulin.
Synthetic genes for human insulin A and B chains were cloned separately in plasmid pBR322. The cloned synthetic genes were then fused to an Escherichia coli beta-galactosidase gene to provide efficient transcription and translation and a stable precursor protein. The insulin peptides were cleaved from beta-galactosidase, detected by radioimmunoassay, and purified. Complete purification of the A chain and partial purification of the B chain were achieved. These products were mixed, reduced, and reoxidized. The presence of insulin was detected by radioimmunoassay.
D000596:Amino Acids / Q000032:analysis; D004274:DNA, Recombinant; D000939:Epitopes; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D008040:Genetic Linkage; D006801:Humans; D007328:Insulin / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D046911:Macromolecular Substances; D010957:Plasmids; D001616:beta-Galactosidase / Q000235:genetics
D000596:Amino Acids; D004274:DNA, Recombinant; D000939:Epitopes; D007328:Insulin; D046911:Macromolecular Substances; D001616:beta-Galactosidase