<p align="left" style="background-color:#ffffff;"> <br><br><br>Team Plant Promoter consisted of Ryan and Niall. Their objective was to isolate four plant promoters that are found in <i> Arabidopsis thaliana </i> and create gene constructs with either luciferase or our TSH antagonist as the coding sequence. The plant promoters were isolated by grinding up <i> Arabidopsis thaliana </i> tissue, extracting the DNA, and amplifying the promoter sequences using PCR, with primers that would incorporate the BsmB1 type IIS restriction endonuclease recognition site, and only amplify the 3'-most 500bp of promoter DNA. These were then excised from a gel and purified. The DNA was put into the level 0 plasmid and can be seen on the <a href="https://2017.igem.org/Team:Cardiff_Wales/basicparts">basic parts</a> page. These promoters are PDF1.2 (jasmonic acid inducible), PR2 and GST6 (salicylic acid inducible), and WRKY30 (DAMP response inducible). These were then used to create level 1 constructs containing Luc+ or our TSH antagonist with his tags. Unfortunately, the promoter:luc+:NosT constructs grew in <i> E. coli </i>, but not in <i> Agrobacterium </i> when we ran out of lab time, so these constructs could not be tested. WRKY30 constructs with our TSH antagonist did not grow either. However, the TSH-antagonist <i> Agrobacterium</i>-constructs with PDF1.2, PR2, and GST6 were produced, and can be seen on our <a href="https://2017.igem.org/Team:Cardiff_Wales/compositeparts"> composite parts</a> page. These were used to transform our <i> Nicotiana benthamiana </i> plants, and had protein extractions performed on them following induction with the associated acid for ~6 hours. Sadly, these gels were not conclusive, with no bands present definitively in the samples with nickel beads to isolate the protein. Regardless, these gels can be seen on our <a href="https://2017.igem.org/Team:Cardiff_Wales/results"> Results </a> page.