Abstract
Background: Human epidermal growth factor (hEGF) has drawn intense research attention due to its potential ability to promote healing of serious injuries, such as cuts, burns, and diabetic ulcers. Although hEGF displays prospective clinical value, the growth factor is restricted to the treatment of chronic diabetic ulcers because of its high production cost. Methods: Leguminous plant peanut (Arachis hypogaea L.) hairy roots contain relatively few toxic and harmful substances, and tested as an excellent production system for hEGF in our study. To explore the possibility of hEGF expression in peanut, hEGF overexpression hairy roots were obtained by infecting leaves with Agrobacterium rhizogenes R1601. Results: The maximum transgenic hairy roots inducing rate was 82%. Protein purification and mass spectrometry assays showed that the protein expressed in peanut hairy roots was identified as hEGF. Furthermore, Methylthiazolyldiphenyl-tetrazolium bromide assay showed that hEGF promoted HL-7702 liver cells proliferation, which indicate that hEGF has biological activity and non-toxic on human cells. Conclusion: Our results demonstrate the capacity of peanut hairy root cultures as a controlled, sustainable, and scalable production system that can be induced to produce valued human proteins, such as hEGF.
Highlights
Human epidermal growth factor, a 53-amino acid peptide with three disulfide bonds, promotes the proliferation and differentiation of epithelial cells [1,2,3]. hEGF is specially recognized by membrane-localized receptor, hEGF receptor (EGFR) in human and initiates a series of intracellular signalling cascades, including the synthesis of proteins, the improvement of DNA topoisomerase activity and the activation of key genes involved in cell proliferation, differentiation, and migration [4,5]
To establish the protein expression system in peanut hairy root, the green fluorescent protein (GFP) gene was constructed into the pCAMBIA1300 expression vector, which was used as the experimental vector to explore and optimize the induction conditions of peanut hairy roots
The plasmid was subsequently transformed into Agrobacterium rhizogenes R1601, and the strain carrying hEGF was designated as HR1601
Summary
Human epidermal growth factor (hEGF), a 53-amino acid peptide with three disulfide bonds, promotes the proliferation and differentiation of epithelial cells [1,2,3]. hEGF is specially recognized by membrane-localized receptor, hEGF receptor (EGFR) in human and initiates a series of intracellular signalling cascades, including the synthesis of proteins, the improvement of DNA topoisomerase activity and the activation of key genes involved in cell proliferation, differentiation, and migration [4,5]. Human epidermal growth factor (hEGF), a 53-amino acid peptide with three disulfide bonds, promotes the proliferation and differentiation of epithelial cells [1,2,3]. Owing to the high production cost of hEGF, treatment with the growth factor is limited to chronic diabetic ulcers. HEGF displays prospective clinical value, the growth factor is restricted to the treatment of chronic diabetic ulcers because of its high production cost. Protein purification and mass spectrometry assays showed that the protein expressed in peanut hairy roots was identified as hEGF. Conclusion: Our results demonstrate the capacity of peanut hairy root cultures as a controlled, sustainable, and scalable production system that can be induced to produce valued human proteins, such as hEGF
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