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Recombinant Human Insulin-Like Growth Factor II Expressed in Escherichia coli

Bio/Technology volume 5, pages 1047–1051 (1987)Cite this article

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.

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Author information

Author notes

  1. George L. Long

    Present address: Department of Biochemistry, School of Medicine, Given Building, University of Vermont, Burlington, VT, 05405

  2. Michele C. Smith: To whom correspondence and reprint requests should be sent.

Authors and Affiliations

  1. Department of Biochemistry, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, 46285

    Thomas C. Furman & Michele C. Smith

  2. Department of Molecular Genetics, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, 46285

    Janet Epp & Brigitte Schoner

  3. Department of Molecular Biology, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, 46285

    Hansen M. Hsiung, JoAnn Hoskins, George L. Long & Dennis P. Smith

  4. Department of Central Nervous System Research, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, 46285

    Laurane G. Mendelsohn

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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

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