Promoters

RD29A Promoter + Strong Plant Kozak (RBS) + RFP Team:Nevada/registry submissions

The 2010 Nevada iGEM team has three goals for this year’s competition. First, we are going to test the validity of utilizing Nicotiana tabacum protoplasts (NT cells), plant cells without the cell wall, as a model for the expression of higher plant genes for future iGEM competitions. This system is useful in the respect that the time it takes to obtain transgenic lines of cells is greatly reduced compared to the time to obtain transgenic plants. These cells can therefore be utilized as a quick proof-of-concept test model before moving synthetic constructs into plants of interest. We also aim to produce an iGEM-compatible plant-specific plasmid, several stress-inducible plant promoters, reporter genes containing Kozak sequences (ribosome binding sites) and terminators that conform to BioBrick standards. Lastly, we hope to measure the induction of these stress promoters in real-time by performing a fluorometry assay in which stress will be applied to NT cells and fluorescent output by a reporter (GFP) will be measured to detail induction in real time. This method has a distinct advantage over microarrays since microarrays are only one ‘snapshot’ in time.