Team:Lethbridge/Parts
In order to explore the modularity and various applications of Protein Nanocompartments (PNCs), we have designed systems that have distinct PNCs, surface modifiers, and cargos. Most PNCs are designed to have a specific surface modifying feature and cargo that look at different encapsulation strategies that are specific to these specific PNCs.
Each PNC system covers distinct applications for Research use. Each of our PNC systems is described in detail on our design page.
There are several capsid proteins described in the registry, including the MS2 protein. Our project diversified the capsid proteins available on the registry and are designed as composite parts under control of a T7 inducible promoter (BBa_I712074) a medium strength RBS (BBa_J61100) and a double terminator (BBa_B0014).
Parts Submitted:
This year, we submitted a pSB1C3 vector containing the "minimal Arc Gag" (derived from HIV-1 homology) construct to the registry. This construct was theorized and modeled by our team this year. The Arc DNA sequence, originally from Rat was compared to the HIV-1 DNA sequence due to Arc's relationship to retrotransposons. This part is theorized to retain its abilities to form capsid structures and deliver RNA to cells. Our results can be found here. We hope to finish cloning all of our designed parts for future studies and applications.
Basic and Composite Parts:
All of our new parts can be found on the basic parts page and have been placed on the iGEM registry. These contain the four protein nanocompartments and the various cargo proteins and external surface modifiers.
This year we have designed 9 composite parts consisting of 4 capsid proteins with surface modifiers, cargoes with specific modifications for encapsulation and an external part for the surface modification of the P22 capsid. Each composite part has the same design, consisting of the T7 inducible promoter (BBa_I712074), a medium strength RBS (BBa_J61100) and a double terminator (BBa_B0014).
Improved Parts:
As our system deals with the encapsulation of specific cagos one of our future applications is the encapsulation of Cell Free systems. Therefore we are pleased to say that our continuing work with last years project, Next Vivo, will allow us to encapsulate our cell free system. Since last years Jamboree we have been able to clone and characterize 15 more parts and will now submit them to the registry as improved parts. These parts include a his tag on the N or C terminus for purification and follow the general design of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and Double terminator (BBa_B0015).
| Biobrick Number | Part Name |
|---|---|
| BBa_K2683016 | Release Factor 1 |
| BBA_K2683017 | Release Factor 2 |
| BBa_K2683018 | Cysteine tRNS Synthetase (CysRS) |
| BBa_K2683019 | Lysine tRNA Synthetase (LysRS) |
| BBa_K2683020 | Phenylalanine tRNA Synthetase alpha subunit (PheRS alpha) |
| BBa_K2683021 | Serine tRNA Synthetase (SerRS) |
| BBa_K2683022 | Tryptophan tRNA Synthetase (TrpRS) |
| BBa_K2683023 | Valine tRNA Synthetase (ValRS) |
| BBa_K2683024 | Inititation Factor 1 |
| BBa_K2683025 | Initiation Factor 3 |
| BBa_K2683026 | Elongation Factor-Thermo Unstable (EF-Tu) |
| BBa_K2683027 | Elongation Factor- Thermo Stable (EF-Ts) |
| BBa_K2683028 | MyoKinase (MK) |
| BBa_K2683029 | Nucleoside Diphosphate Kinase (NDK) |
| BBa_K2683030 | Peptidyl Prolyl Isomerase (PPiase) |
Existing Parts on the Registry
For the design of our protein nanocompartments, we have utilized previous parts already found in the registry:
| Biobrick Number | Part |
|---|---|
| BBa_I712074 | T7 Promoter |
| BBa_J61100 | Medium strength ribosomal binding site |
| BBa_B0014 | Double Terminator |