Recombinant Human Insulin-Like Growth Factor II Expressed in Escherichia coli
Author: ["Thomas C. Furman","Janet Epp","Hansen M. Hsiung","JoAnn Hoskins","George L. Long","Laurane G. Mendelsohn","Brigitte Schoner","Dennis P. Smith","Michele C. Smith"]
Publication: Bio/Technology
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Abstract
Human Insulin-like growth factor II (IGF-II) was produced in Escherichia coli as a 113 amino acid chimeric protein, LE1-Met-IGF-II consisting of 45 N-terminal amino acids from the E. coli trp leader peptide and the trpE polypeptide, and 67 C-terminal amino acids for IGF-II. High-level expression of the fusion protein, LE1-Met-IGF-II, was obtained with pCZ21, a plasmid containing a thermoinducible runaway replicon and the kanamycin resistance marker from Tn5. In E. coli, LE1-Met-IGF-II formed granules that were isolated and cleaved with cyanogen bromide to liberate IGF-II. IGF-II was sulfitolyzed, purified by anion exchange chromatography, refolded through a disulfide interchange reaction, and further purified by reverse-phase HPLC and gel filtration. The biological activity of this recombinant human IGF-II was measured in a competitive protein binding assay for IGF-II and by its ability to stimulate amino isobutyric acid uptake and protein synthesis in NCTC 2414 fibroblast cultures.
Cite this article
Furman, T., Epp, J., Hsiung, H. et al. Recombinant Human Insulin-Like Growth Factor II Expressed in Escherichia coli. Nat Biotechnol 5, 1047–1051 (1987). https://doi.org/10.1038/nbt1087-1047
