Team:Calgary/Lab/Mutant

Overview
Updates

Circuits

• Signalling Circuit
• Reporter Circuit
• Mutant Circuits
• Response Circuit

Quorum Sensing
Chassis Selection

Protocol & Safety
Notebook & Parts

Figure 1. Schematic diagram of LuxO D47A (left) and LuxO D47E (right) mutant circuits. The circuits are almost identical in that they have the same promoter (BBa_R0040), RBS (BBa_B034) and terminator (BBa_B0015). This design will allow for constitutive expression of these proteins. The LuxO D47A mutant mimics the unphosphorylated and thus inactive form of LuxO, meaning that it will not bind to the qrr4 promoter. The LuxO D47E mutant, however, mimics the phosphorylated and thus active form of LuxO, and will thus bind to the qrr4 promoter and induce expression of downstream genes. Once these circuits are tested with a non-Biobrick reporter (in the KT1144 cells) they are used to test whether the reporter circuit is functional. If in the presence of the LuxO D47A mutant the reporter cells do not glow and if in the presence of the LuxO D47E mutant the reporter cells do glow, we know the reporter circuit is functional. Purpose: To test the functionality of the Reporter and Response circuits. LuxO D47A mimics the unphosphorylated and thus inactive form of LuxO, whereas LuxO D47E mimics the phosphorylated and thus active form of LuxO.