Team:NJMU-China/Engineering
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Overview
We have successfully completed the synthesis of our parts. Also, we simulated the interactions and verified success of our engineering preliminarily.
Success in Constructing Parts
In the first place, we have successfully optimized the color-reaction reporting system (Figure 1).
Figure 1. Color-Reaction of pGEX-6P-1
Then, we have successfully constructed all the parts and sequenced them for verification.
Figure 2. Enzyme Digestion Result of Serotonin Biosensor Version 2
Demonstration of Feasibility about Our Design
We successfully constructed the 3D structure of proteins (LasR, LacZ), DNA fragments (PlasI) and small molecules (Serotonin, Dopamine, 3-O-C12-HSL, X-gal) involved in this project (See more in Simulations of Interaction ).
We evaluated all interactions involved in our project by calculating docking scores. The results show that, the docking of LasR-Serotonin/Dopamine ~ PlasI have scores of -260.09 and -258.87 (the greater the absolute value, the more informative) (Table 1). We also computed the docking score of LasR-HSL ~ PlasI (Positive Control) and LasR ~ PlasI (Negative Control). We found that the docking of LasR-Serotonin/Dopamine ~ PlasI have almost same score with the docking of LasR-HSL ~ PlasI at similar ligand RMSD, while having apparently higher score in comparison with the docking of LasR ~ PlasI.
In summary, the protein complex of LasR-Serotonin has similar performance with the experimental verified LasR-HSL complex in docking with PlasI, indicating its feasibility to be designed into a biosensor.
| Interaction Type | Docking score | Ligand rmsd (Å) |
| LasR-HSL/PlasI | -257.85 | 135.92 |
| LasR-serotonin/PlasI | -260.09 | 135.53 |
| LasR-dopamine/PlasI | -258.87 | 167.30 |
| LasR/PlasI | -229.76 | 174.41 |
Table 1. Docking Scores of Different Interactions