Results and implications for Iowa iGEM 3HP biobrick construct

Our biobrick is designed to luminesce, or emit off light, in the presence of 3HP. Induction of luminescence by 3HP is seen in Figure 2. However, this only occurs when cells have been cultured in the presence of 3HP for 18 hours, and the growth curve in Figure 1 displays a dramatic lag in growth at the highest concentrations of 3HP (greater than 30mM). Furthermore, when compared to our empty vector in Figure 3 there is no difference in induction. This raises significant questions about the specificity of our reporter. Finally, in these experiments we included similar previously developed constructs to compare the sensitivity of the two reporters.¹ Successful induction with their construct evidenced that the assay itself worked and that their construct has superior sensitivity. More troubleshooting is needed to optimize the performance and design of our construct.

1. Erik K. R. Hanko, Nigel P. Minton, and Naglis Malys. "Characterisation of a 3-hydroxypropionic Acid-inducible System from Pseudomonas Putida for Orthogonal Gene Expression Control in Escherichia Coli and Cupriavidus Necator." Scientific Reports 7, no. 1 (2017): 1724.