Benth Biofactory
The Cardiff iGEM team of 2017 aimed to produce a high level of gene expression from four promoter constructs in the Nicotiana benthamiana expression system. These constructs were put together using golden gate ligation. Our promoters respond to several different stimuli, are native to Arabidopsis thaliana, and have been characterized previously. We aimed to both quantify their expression levels using luciferase transcriptional units, and use them to create a thyroid stimulating hormone (and thyroid stimulating immunoglobulin) antagonist that could be used to treat Graves' disease. Consequently, we have added several parts to the phytobrick registry. Our antagonist, called TSHantag, has not been previously used as a therapeutic agent, and could be tested in vitro if purified. More details about the project can be found on our project description page.
We have expertise using the tobacco expression system and so welcomed collaborations with iGEM teams who wish to test the function of their proteins in plants!
Project Abstract
Grave’s Disease arises from the overproduction of the thyroid stimulating hormone (TSH). Such overproduction leads to increased thyroxine levels, consequently resulting in hyperthyroidism. Our project will involve the expression of the human thyroid stimulating hormone antagonist (TSHantag) using the Tobacco expression system. As of yet, there have been no examples using the tobacco system for expression of the TSHantag protein. Therefore, we will design and generate unique transcriptional units (TUs) for expression of TSHantag in tobacco. These TUs will be introduced into tobacco using agrobacterium-mediated transformation. We will optimise the amounts of heterologous protein that can be produced in this system before collaborating with researchers at University Hospital of Wales to test the efficacy of the TSHantag in an in vitro system.
