Our contributions include a number of gene fragments that we designed using sequences derived from published journal articles with the overall goal to create a sensitive guanine biosensor. These gene fragments were designed to fit the MoClo GoldenGate cloning scheme for ease of use in any projects using the MoClo system. We worked on two separate genetic approaches to accomplish the same goal: the goal - “Create a bacteria that is fine tuned to produce reporter pigment in response to specific amounts of guanine produced by varroa mites.”

In an effort to achieve this, we crafted two genetic approaches: the Guanine-II Riboswitch and the purR Purine Sensor. For more information, consult our Description Tab.

We created Level 0 Golden Gate parts including promoters, ribosome binding site, coding sequences, and terminators. We also created Level 1 Golden Gate parts that are a synthesis of the Level 0 Golden Gate parts that in combination act as experimental guanine biosensors. For more information on these parts, please visit our engineering page. Parts were codon optimized for E. coli and designed to be Golden Gate compatible with Bsa1 sites for cloning into circular plasmids.

Our project after designing the gene fragments involved cloning these parts together and characterizing the plasmid products as biosensors in response to guanine.