Part:BBa_K4190017

GLP-1 Receptor Gene Insert Optimized for E. coli

Protein coding region for expressing GLP-1 R in E. coli.

Usage and Biology

The native GLP1 receptor (GLP1-R) is found on the surface membrane of pancreatic cells and is 463 amino acids in size [1]. The binding domain of the receptor is 365 base pairs and feasible for production in a recombinant vector. The crystal structure of binding between Ex-4 and the GLP1-R allows for a truncated GLP1-R (GLP1-RΔ) to be an effective alternative for testing of binding in the respective binding pocket [2]. The GLP1-R gene block of 505 bases will be inserted into the pET28 backbone via golden gate and expressed in BL21 E. coli cells. The GLP1-R protein will be purified for use during Ex-4 binding affinity tests. A binding affinity test between the truncated GLP1-R and Ex-4 via an octet machine will allow for the secondary folding of the Ex-4 and functionality to be qualitatively deduced. The binding affinity of manufactured Ex-4 is very strong at 6 nmol [3].

Sequences and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] “Glucagon Like Peptide 1 Receptor - an overview | ScienceDirect Topics.” https://www.sciencedirect.com/topics/medicine-and-dentistry/glucagonlike- peptide-1-receptor/ (accessed Aug. 28, 2022).

[2] “GLP1R glucagon like peptide 1 receptor [Homo sapiens (human)] - Gene - NCBI.” https://www.ncbi.nlm.nih.gov/gene/2740 (accessed Aug. 28, 2022).

[3] M. K. K. Yap and N. Misuan, “Exendin-4 from Heloderma suspectum venom: From discovery to its latest application as type II diabetes combatant,” Basic & Clinical Pharmacology & Toxicology, vol. 124, no. 5, pp. 513–527, 2019, doi: 10.1111/bcpt.13169.