Part:BBa_K2152000
human Epidermal Growth Factor(hEGF) with His-tag
Human EGF（epidermal growth factor）is a 6045-Da protein with 53 amino acid residues. A signal peptide phoA is added at the N-terminus to facilitate secretion. We engineered two peptide tags, a phoA signal peptide at the N-terminus and a His tag at the C-terminus of hEGF protein. PhoA is a fragment of signal peptide that guides the protein to go across cytomembrane and do not decrease activity of the protein. His-tag is an amino acid motif in proteins that consists of at least six histidine (His) residues, at the N-terminus of the EGF protein. It is also known as hexa histidine-tag. hEGF is originally described as a growth factor to stimulate cell growth, proliferation and differentiation by binding to its receptor EGFR. In our project, we used hEGF as a therapeutic agent to treat Inflammatory bowel diseases.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 198
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Figure 1. Western Blot of EGF with his-tag. The red rectangle marked area is our expressed EGF protein. The track on the left is the control group without IPTG induction.
During the competition, we sent our purified EGF expressed by our engineered bacteria to team SYSU-MEDICINE, and they use their IBD mouse model to help us prove its validity.
They referred to the article [1] and designed the following experiment:
Figure 2. IBD mice in the control group almost kept losing weight during these five days. While mice in experiment group even gained weight on the last day.
Figure 3. The length of colons indicates the effect of repair. The longer the colon is, better healing effect it is. They cannot measure the length of colons when mice were alive, but we can still observe the effect of EGF through longer length of colons in the experimental group.
[1]Modeling method reference: Chemically induced mouse models of intestinal inflammation Stefan Wirtz, Clemens Neufert, Benno Weigmann & Markus F Neurath Laboratory of Immunology, I Medical Clinic, University of Mainz, Mainz, Germany. Correspondence should be addressed to M.F.N. Published online 15 March 2007; doi:10.1038/nprot.2007.41
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