During the first steps of the design process, our iGEM team discussed what engineering aims our project wanted to achieve. It was an important first step in determining the goals for our research. We based our engineering aims on Endy's "Foundations for engineering biology," from 2005.
After determining our engineering goals and aims for this year's competition. Our team needed to do background research into quorum sensing to familiarize ourselves with the terminology going to be used for this competition. In addition to this, our team needed to reaffirm the 2016 ASU iGEM team's data collected from last year before going any further with designing our project. While confirming last year's team data, we discovered a major flaw in their design process of the receivers. These receivers were showed poor expression and only a select few were good enough to characterize. In addition to this, the system these receivers were cloned into did not show a good cloning ratio. In fact, it was very difficult to clone these receivers. After this, we realized we need to re-evaluate these receivers to determine whether it was the system we were cloning them in was bad, or the receivers themselves that needed to be redesigned.
From this, it was determined that the receivers themselves were bad and needed to be redesigned this year. From this realization, our design process was re-evaluated and is represented in the flow chart below:
Describe what each of the bio-bricks used during the design. In addition, tell the bio-brick parts of the senders use. Also the picture of the sender vector. Modular center vector on benchling.
| Receiver Part Names | Part Number | Description |
|---|---|---|
| Inducible Promoter | ||
| Ribosomal binding site for GFP | BBa_B0032 | |
| Green Fluorescent Protein | BBa_E0040 | |
| Terminator for Green Fluorescent Protein | BBa_J61048 | |
| Promoter for Receiver Protein | BBa_J23100 | |
| Ribosomal Binding Site for Receiver Protein | BBa_B0034 | |
| Receiver Protein (LasR, RpaR, TraR) | ||
| Terminator for Receiver Protein | BBa_B0015 |
| Sender Part Name | Part Number | Part Type |
|---|---|---|
| C12-HSL, N-(2-oxooxolan-3-yl)dodecanamide Sender-AubI | BBa_K2033000 | Sender |
| Isovaleryl-HSL, 3-methyl-N-[(3S)-2-oxooxolan-3-yl]butanamide Sender-BjaI | BBa_K2033002 | Sender |
| 3-phenyl-HSL, (3S)-3-[(2-oxo-3-phenylpropyl)amino]oxolan-2-one Sender-BraI | BBa_K2033004 | Sender |
| 3OH-7-cis-C14-HSL, (Z)-3-hydroxy-N-[(3S)-2-oxooxolan-3-yl]tetradec-7-enamide Sender-CerI | BBa_K2033006 | Sender |
| C8-HSL, N-[(3S)-2-oxooxolan-3-yl]octanamide Sender*-SinI | BBa_K2033008 | Sender |
[1] Endy, Drew. "Foundations for engineering biology." Nature 438.7067 (2005): 449
