Team:UZurich/Parts
Parts
Due to the novelty of our project's nature and the lack of knowledge and intense research on the topic, we add a solid foundation of 8 basic parts and 6 composite parts to the registry. To increase the usability of our parts for others, we have characterized our parts in as much detail as possible with text, plots and figures.
Our parts fall into three categories:
1. Display of plant immune response elicitors on OMV surface
The main goal of our project is to use and engineer OMVs to protect plants from pathogens. To achieve this goal, the following parts were used to enhance the protective immune responses triggered by OMVs. Among them, we have elf18 (BBa_K3989023) as our Best Basic Part, and ClyA-elf18 (BBa_K3989011) as our Best Composite Part.
| Description | Part Number | Type | Designer |
|---|---|---|---|
| elf18 | BBa_K3989023 | Basic | Songyuan Zhang, Tianyu Xu |
| ClyA-flg22 | BBa_K3989010 | Composite | Songyuan Zhang, Tianyu Xu |
| ClyA-elf18 | BBa_K3989011 | Composite | Songyuan Zhang, Tianyu Xu |
| ClyA-elf18-flg22 | BBa_K3989013 | Composite | Songyuan Zhang, Tianyu Xu |
| ClyA-dCBD | BBa_K3989014 | Composite | Songyuan Zhang, Tianyu Xu |
2. EsaR-based quorum-sensing system
Unlike the classic quorum-sensing regulator LuxR which can only upregulate gene expression in response to QS molecules, the EsaR regulator can work as either an activator or a repressor, depending on the promoter used. Here we introduced three EsaR variants with different sensitivities to 3OC6HSL, as well as activatable and repressible promoters. In our project, we use them to construct an inducible OMV production system by downregulating TolB expression and by upregulating TEV protease, simultaneously (see our Design page). We hope they can help future teams to construct better quorum-sensing systems.
| Description | Part Number | Type | Designer |
|---|---|---|---|
| EsaR I70V mutant | BBa_K3989003 | Basic | Tianyu Xu, Songyuan Zhang |
| EsaR D91G mutant | BBa_K3989004 | Basic | Tianyu Xu, Songyuan Zhang |
| EsaR V220A mutant | BBa_K3989005 | Basic | Tianyu Xu, Songyuan Zhang |
| PesaR-C | BBa_K3989008 | Basic | Tianyu Xu, Songyuan Zhang |
| PesaS | BBa_K3989009 | Basic | Tianyu Xu, Songyuan Zhang |
| EsaR PesaR-C GFP | BBa_K3989024 | Composite | Tianyu Xu, Songyuan Zhang |
| EsaR PesaS GFP | BBa_K3989025 | Composite | Tianyu Xu, Songyuan Zhang |
For a detailed characterization, please refer to each part via the links in the tables below as well as the improvement section on our Results page.
3. TEV protease variants with improved efficiency
Here we introduced some TEV protease variants with enhanced efficiency, which were generated by directed evolution from two newly published papers [1][2]. In our project, they were used to construct an inducible OMV production system (see our Design page). Considering that TEV protease is also useful in many other scenarios, for instance, protein purification (in vitro) and synthetic signaling pathways (in vivo), we hope that introducing these TEV protease variants to the Registry can benefit subsequent iGEM teams a lot.
| Description | Part Number | Type | Designer |
|---|---|---|---|
| uTEV3 | BBa_K3989001 | Basic | Tianyu Xu, Songyuan Zhang |
| eTEV | BBa_K3989002 | Basic | Tianyu Xu, Songyuan Zhang |
[1] Sanchez, M. I., & Ting, A. Y. (2020). Directed evolution improves the catalytic efficiency of TEV protease. Nature methods, 17(2), 167-174.
[2] Denard, C. A., Paresi, C., Yaghi, R., McGinnis, N., Bennett, Z., Yi, L., ... & Iverson, B. L. (2021). YESS 2.0, a Tunable Platform for Enzyme Evolution, Yields Highly Active TEV Protease Variants. ACS Synthetic Biology, 10(1), 63-71.