Parts:
We successfully construct a plasmid that could secrete turboPETase, and we register the basic components
of the plasmid into igem parts:
| Code | Name | Description |
|---|---|---|
| BBa_K5233025 | TurboPETase(LamB) | A designed enzyme that can be secreted into the extracellular space |
| BBa_K5233021 | TurboPETase(MalE) | A designed enzyme that can be secreted into the extracellular space |
| BBa_K5233022 | TurboPETase(OmpA) | A designed enzyme that can be secreted into the extracellular space |
| BBa_K5233023 | TurboPETase(PelB) | A designed enzyme that can be secreted into the extracellular space |
Among the 4 kinds of TurboPETase, TurboPETase(OmpA) has the best degradation effect. Thus we choose it as out best new best part for our gold medal.
We also designed a TPA transporting system plasmid composed of the following basic parts:
| Code | Name | Description |
|---|---|---|
| BBa_K5233006 | tpiA | A kind of tripartite tricarboxylate transporter(TTT) family protein |
| BBa_K5233007 | tpiB | A kind of tripartite tricarboxylate transporter(TTT) family protein |
| BBa_K5233008 | tphC | tphC, a kind of substrate-binding protein (SBP) belonging to the tripartite tricarboxylate transporters |
We composite these parts to form a composite part transport system:
| Code | Name | Description |
|---|---|---|
| BBa_K5233044 | tpiA-tpiB-tphC | A transporter system in C.elegans, a system that can transport TPA into the C.elegans intestinal cell |
Besides, we design a sensor system which responds to the presence of TPA:
| Code | Name | Description |
|---|---|---|
| BBa_K5233009 | tphR | A kind of regulatory protein that can form complex with TPA which will activate PtphC |
| BBa_K5233010 | PtphC | PtphC is a kind of promter that responds to the presence of TPA |
| BBa_K5233011 | eGFP | A kind of green fluorescent protein |
We composite these parts to form a composite part transport system:
| Code | Name | Description |
|---|---|---|
| BBa_K5233030 | tphR-PtphC-eGFP | A biosensor element, an element that can detect the degradation of PET |
Besides, we design a plasmid that could secrete MHETase to promote the degradation of PET:
| Code | Name | Description |
|---|---|---|
| BBa_K5233012 | MHETase(Amcut) | A designed enzyme that can be secreted into the extracellular space and degrade MHET |
| BBa_K5233013 | MHETase(pelB) | A designed enzyme that can be secreted into the extracellular space and degrade MHET |
In addition, we try to express transporter system in the intestinal cell of C.elegans and we register some new basic parts with specific functions:
| Code | Name | Description |
|---|---|---|
| BBa_K5233000 | vha-6 | A specific promoter that only expressed in the gut of C.elegans |
| BBa_K5233040 | gbb-1 | A kind of signal peptide that can guide protein locate on Intestinal cell membrane, it is used for protein that has 7 transmembrane domains |
| BBa_K5233050 | glr-2 | A kind of signal peptide that can guide protein locate on Intestinal cell membrane, it is used for protein that has 3 transmembrane domains |
| BBa_K5233042 | dsl-1 | A kind of signal peptide that can guide protein secretion |
| BBa_K5233043 | SL2 | Spliced leader 2 |
We composite the transport system designed for C.elegans:
| Code | Name | Description |
|---|---|---|
| BBa_K5233032 | vha6-gbb1-tpiA-SL2-glr2-tpiB-SL2-dsl1-tphC | A transporter element, an element that can transport TPA into the cell |
Hardware:
In the process of design and research, we found that there are some inconveniences in the application
of 24-well plates in nematode research related projects. Therefore, we proposed some optimized designs
based on the nematode culture mode and detection mode. We hope to achieve the integration of solid-liquid
culture, separated detection and more convenient nematode handling. We hope that this design can be applied
to nematode culture scenarios in the laboratory to ensure biosafety while promoting the application and
development of nematode-related research.
In addition, in order to link the laboratory scenario with industrial treatment, we designed a C. elegans-E. coli
co-culture fermenter to degrade microplastic wastewater through E. coli secretion of enzymes and primary enrichment,
as well as secondary enrichment of E. coli by nematodes to ultimately achieve biological remediation.