Skip to main content
Log in

Expression of recombinant epidermal growth factor inE. coli

  • Published:
Biotechnology and Bioprocess Engineering Aims and scope Submit manuscript

Abstract

Epidermal growth factor (EGF) known as a urgastrone is a powerful mitogen with a wide variety of possibilities for medical usages. A mature EGF coding region was isolated from human prepro-EGF sequence by a conventional PCR and cloned into pQE vector in which the gene product was supposed to be expressed with 6×His tag for the subsequent purification. The recombinant mature EGF was expressed in M15[Rep4], anEscherichia coli host strain, in amount of 30–40% of total proteins present inE. coli extract by the addition of isopropylthio-β-galactopyranoside (IPTG). The recombinant EGF purified using a Ni2+-NTA affinity column chromatography was active in its ability to induce phosphorylation on tyrosine residues of several substrate proteins when murine NIH3T3 and human MRC-5 fibroblast cells were stimulated with it. This work may provide the basic technology and information for the production of recombinant EGF.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Lehinger, A. L., D. L. Nelson, and M. M. Cox (1993) Principles of Biochemistry, 2nd edit, p. 775, Worth Publishers, New York, NY.

    Google Scholar 

  2. Cooke, R. M., A. J. Wilkinson, M. Baron, A. Pastore, M. J. Tappin, I. D. Campbell, H. Gregory, and B. Sheard (1987) The solution structure of human epidermal growth factor.Nature 327: 339–341.

    Article  CAS  Google Scholar 

  3. Hudziak, R. M., J. Schlessinger, and A. Ulrich (1987) Increased expression of the putative growth factor receptor p185HER2 causes transformation and tumorigenesis of NIH3T3 cells.Proc Natl. Acad. Sci. USA 84: 7159–7163.

    Article  CAS  Google Scholar 

  4. Velu, T. J., L. Beguinot, W. C. Vass, M. C. Willingham, G. T. Merlino, I. Pastan, and D. R. Lowry (1987) Epidermal Growth factor-Dependent Transformation by a Human EGF Receptor Proto-Oncogene.Science 238: 1408–1410.

    Article  CAS  Google Scholar 

  5. Heidaran, M. A., T. P. Fleming, D. P. Bottaro, G. I. Bell, P. P. Di Fiore, and S. A. Aaronson (1990) Transformation of NIH3T3 fibroblasts by an expression vector for the human epidermal growth factor precursor.Oncogene 5: 1265–1270.

    CAS  Google Scholar 

  6. The QIAexpressionist (1992) QIAexpress: the high level expression & protein purification system.

  7. Aaronson, S. A. and S. R. Tronick (1991) Growth factor signalling pathways and their alterations in human tumors.FORUM: 14–29.

  8. Kraus, M. H., P. Fedi, V. Starks, R. Muraro, and S. A. Aaronson (1993) Demonstration of ligand-dependent signalling by the erbB-3 tyrosine kinase and its constitutive activation in human breast tumor cells.Pro. Natl. Acad. Sci. USA 90: 2900–2904.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Il Yup Chung.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yoon, C.S., Lee, E.G., Lee, Y.S. et al. Expression of recombinant epidermal growth factor inE. coli . Biotechnol. Bioprocess Eng. 2, 86–89 (1997). https://doi.org/10.1007/BF02932330

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02932330

Key words

Navigation