title: Expression in Escherichia coli of a chemically synthesized gene for a "mini-C" analog of human proinsulin.
abstract: A gene has been constructed which codes for an analog of human proinsulin in which the normal 35-amino acid connecting peptide is replaced by a "mini-C" peptide of six amino acids (Arg-Arg-Gly-Ser-Lys-Arg). The gene, composed of oligonucleotide fragments synthesized by the triester method, was cloned and expressed as a beta-galactosidase hybrid protein. The proinsulin analog was separated from beta-galactosidase by cyanogen bromide cleavage and purified. Controlled disulfide exchange in the S-sulfonate of the analog generated a molecule having high-pressure liquid chromatography (HPLC) and radioimmunoassay (RIA) behavior consistent with a proinsulin-like structure.
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D003488:Cyanogen Bromide; D004926:Escherichia coli / Q000235:genetics; D005813:Genes, Synthetic; D007328:Insulin / Q000032:analysis; D009842:Oligopeptides / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D010957:Plasmids; D011487:Protein Conformation; D011863:Radioimmunoassay
chemical_list: D007328:Insulin; D009842:Oligopeptides; D003488:Cyanogen Bromide