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Expression of Recombinant Human Epidermal Growth Factor in Escherichia coli and Characterization of its Biological Activity

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Abstract

Recombinant human epidermal growth factor (EGF) was successfully expressed as a fusion protein in Escherichia coli system. This system was used OmpA signal sequence to produce soluble protein into the periplasm of E. coli. Human EGF (hEGF) synthesized in bacterial cell was found to be similar in size with the original protein and molecular weight approximately at 6.8 kDa. Cell proliferation assay was conducted to characterize the biological activity of hEGF on human dermal fibroblasts. The synthesized hEGF was found to be functional as compared with authentic hEGF in stimulating cell proliferation and promoting growth of cell. In comparison of biological activity between synthesized and commercial hEGF on cell proliferation, the results showed there was no significant different. This finding indicates the synthesized hEGF in E. coli system is fully bioactive in vitro.

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Abbreviations

hEGF:

human epidermal growth factor

HDF:

human dermal fibroblasts

IPTG:

isopropylthiogalactopyranoside

EDTA:

ethylenediaminetetraacetic acid

LB:

Luria Bertani

TBS:

tris buffer saline

NaCl:

sodium chloride

Tris-HCl:

tris-hydrochloride

MTT:

methylthiazolyldiphenyl-tetrazolium bromide

E. coli :

Escherichia coli

PBS:

phosphate buffer saline

CO2 :

carbon dioxide

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Acknowledgements

This research was supported by a grant from the Ministry of Science, Technology and Innovation (MOSTI), Malaysia. Grant. no: 03-02-04-0562- SR0008/05-03.

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Authors and Affiliations

  1. Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    Ahmad Faizal Abdull Razis

  2. Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    Elysha Nur Ismail, Zarida Hambali & Muhammad Nazrul Hakim Abdullah

  3. Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    Abdul Manaf Ali

  4. Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    Mohd Azmi Mohd Lila

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  1. Ahmad Faizal Abdull Razis
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  2. Elysha Nur Ismail
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  4. Muhammad Nazrul Hakim Abdullah
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Correspondence to Ahmad Faizal Abdull Razis.

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Abdull Razis, A.F., Ismail, E.N., Hambali, Z. et al. Expression of Recombinant Human Epidermal Growth Factor in Escherichia coli and Characterization of its Biological Activity. Appl Biochem Biotechnol 144, 249–261 (2008). https://doi.org/10.1007/s12010-007-8019-9

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  • DOI: https://doi.org/10.1007/s12010-007-8019-9

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