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<h3>3.Expression of TSHH in tobacco leaves</h3><br> | <h3>3.Expression of TSHH in tobacco leaves</h3><br> | ||
<p>We successful generated a range of level 1 phytobricks that included a plant promotor hooked up to TSHH. We attempted to use these constructs to express TSHH in tobacco leaves in a collaboration between <a href="https://2017.igem.org/Team:Cardiff_Wales/Team_TSH">Team TSH</a> and <a href="https://2017.igem.org/Team:Cardiff_Wales/Team_PlantP">Team Plant_P</a>.<br> | <p>We successful generated a range of level 1 phytobricks that included a plant promotor hooked up to TSHH. We attempted to use these constructs to express TSHH in tobacco leaves in a collaboration between <a href="https://2017.igem.org/Team:Cardiff_Wales/Team_TSH">Team TSH</a> and <a href="https://2017.igem.org/Team:Cardiff_Wales/Team_PlantP">Team Plant_P</a>.<br> | ||
− | Constructs were introduced into tobacco leaves and after | + | Constructs were introduced into tobacco leaves and after 2 days either salicyclic acid (GST/PR2) or jasmonic acid (PDF1.2) was also inflitrated into the leaves in order to induce gene expression. We performed crude protein extracts from these leaves and then compared to agrofiltrated leaves that had not been treated with the appropriate plant hormone. We used <a href="http://www.stratech.co.uk/products/P9100-65-USB">nickel beads</a> to isolate the His-tag TSH protein and compared those samples to crude extracts in an attempt to identify bands that are specfic for the TSHH.<br><br> |
<img src="https://static.igem.org/mediawiki/2017/2/27/T--Cardiff_Wales--GSTPRProteingel.png"/><br><br> | <img src="https://static.igem.org/mediawiki/2017/2/27/T--Cardiff_Wales--GSTPRProteingel.png"/><br><br> | ||
− | <p> This protein gel showing the results of our protein extracts from leaves infiltrated with either GST:TSHH or PR:TSHH constructs. Following construct infiltration, they were then infiltrated with salicylic acid. A protein extract was carried out on the leaves to assess whether or not our TSHH antagonist was present. By comparing with the negative control, we could | + | <p> This protein gel showing the results of our protein extracts from leaves infiltrated with either GST:TSHH or PR:TSHH constructs. Following construct infiltration, they were then infiltrated with salicylic acid. A protein extract was carried out on the leaves to assess whether or not our TSHH antagonist was present. By comparing with the negative control, we could identify any specific bands corresponding to the TSHH protein. In further experiments we will attempt to optimise this expression and purification. <br><br> |
<img src="https://static.igem.org/mediawiki/2017/2/20/T--Cardiff_Wales--PDFTSHHproteingel.png"/><br><br> | <img src="https://static.igem.org/mediawiki/2017/2/20/T--Cardiff_Wales--PDFTSHHproteingel.png"/><br><br> | ||
<p> This protein gel shows the results of infiltrating PDF1.2:TSHH constructs, followed by infiltration of jasmonic acid. The lack of an obvious band in the 'beads' samples, and the fact there is no band unique to infiltrated leaves when compared to the negative control suggests that this experiment did not work as hoped. | <p> This protein gel shows the results of infiltrating PDF1.2:TSHH constructs, followed by infiltration of jasmonic acid. The lack of an obvious band in the 'beads' samples, and the fact there is no band unique to infiltrated leaves when compared to the negative control suggests that this experiment did not work as hoped. | ||
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<h3>5.Public Interaction and Human Practices</h3><br> | <h3>5.Public Interaction and Human Practices</h3><br> | ||
− | <p>We conducted a <a href="https://2017.igem.org/Team:Cardiff_Wales/Survey">survey</a> for almost 300 participants that assessed public opinion on the use of genetic modification for the production of | + | <p>We conducted a <a href="https://2017.igem.org/Team:Cardiff_Wales/Survey">survey</a> for almost 300 participants that assessed public opinion on the use of genetic modification for the production of pharmaceuticals in animals or plants.<br> We also discussed with a range of experts the scientific and economic advantages of using plants for protein production. This allowed us to perform an analysis on the benefits of the plant expression system, which integrated with our modelling work. This can be found on <a href="https://2017.igem.org/Team:Cardiff_Wales/Our_research">Our Research</a> page.<br><br> |
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Latest revision as of 17:02, 31 October 2017