Contribution
Suicide pathway
We performed 3D modeling of the cleaved protein, which also provided more information about the components, (BBa_K4253006) and made some amino acid changes to meet our needs for some "timed death". We then used blue light to regulate the expression of the gene, demonstrating that the EL222 light control system can also regulate the expression of the gene very well.
(Figure1: Blue light switch can regulate the expression of cleaved proteins well)
Escape route
We have provided a new original for the parts list - CheZ (BBa_K4253005).
The motility of E. coli is powered by the rapid rotation of the propeller-like flagellum. The flagellar motor can rotate clockwise or counterclockwise. Counterclockwise rotation produces an oriented/linear swim of the bacteria, while clockwise rotation causes the cells to adjust to a new direction and roll over in place.
CheY is a key regulator of flagellar motility response. When it is phosphorylated, it leads to clockwise flagellar rotation, and the phosphoprotein phosphatase CheZ dephosphorylates CheY, leading to counterclockwise flagellar rotation [1]. It has been shown that when CheZ is knocked out from the chemotaxis pathway, tumbling in E. coli cells predominates, and overexpression of CheZ will inhibit tumbling in E. coli, allowing faster linear movement of E. coli.We constructed a "light-averse" E. coli by using blue light as a condition to stimulate CheZ expression. When blue light was present, CheZ expression in E. coli was high, causing E. coli to move at high speed, while E. coli rolled over in place due to the predominance of clockwise flagellar movement in the area without blue light. In this way, E. coli was stranded in the region without blue light. In this way, we constructed the ability of E. coli to move in a directional manner.
We can construct "biological patterns" through the distribution of blue light. More importantly, we have given microorganisms the ability to "carry".
Figure2.The mechanism by which E. coli "transport" viruses
Random Peptide Library
We offer a brand new original for the parts list - random petide
This is a fully random library of proteins, and as long as there is a defined protein sequence, the protein sequence that interacts with it can be found quickly by using this random peptide library. We can then perform the corresponding structural analysis of its filtered protein.The screened protein can be compared to an existing biological protein sequence to explore whether the protein will have some effect on that organism, or the screened protein can be modified to form a new original.After sequencing analysis of the random peptide library, we obtained 68,190,232 peptides in total and 3,359,176 peptides after removing redundancy. Among these peptides, peptides of length 20 were the most abundant, accounting for 73.5% of the total number of non-redundant peptides (Fig. 3A). We carried out saturation analyses and found that about 35Gb data was far from covering all the kinds of peptides in our random peptide library (Fig. 3B), indicating that our random peptide library has a high diversity and a broad potential for protein interaction investigations and applications. As you can see, our random peptide library is very well constructed and the library capacity is very large. So we can announce the successful construction of our random peptide library and the addition of a new part
Figure3. statistics of the sequencing data about our random peptide library. (A) Statistics of the peptide number of different lengths; (B) Saturation analyses according to randomly sampling from the sequencing data. The abscissa represents the proportion of random sampling including 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% and 90%, and the ordinate represents the number of peptides detected corresponding to the percentage of sampled sequencing data.
Hardware
In the modeling section, we predicted the locations in the city where our equipment would be placed. And some of our needs were clarified based on some existing experimental data.
1.We plan to place our equipment in the city sewer network, which requires our equipment to be as small as possible to avoid blocking the sewer pipes.
2.We need to pre-treat the sewage to filter out the larger solids may clog our pipes or something will interfere with our testing.
3.We need to concentrate the virus upfront to get the concentration of our assay solution to the linear range of the cell sensor.
4.Our equipment needs to be tightly enclosed to prevent our engineered bacteria from leaking into the effluent.
From this, we designed our hardware. Our hardware consists of a pre-treatment chamber, an enrichment chamber, an assay chamber and an overall operating system.
First, we will pre-treat the wastewater with a regular sampling. This is a device similar to a pumping toilet, and the pump will pump water at regular intervals for testing. In the pre-treatment chamber, we simply remove the larger solids with some membranes.
When the liquid level reaches a certain height, the effluent will enter the enrichment chamber for enrichment.
In the enrichment chamber, we will use our "couriers" to deliver viruses as cargo to areas without blue light. This "repelling" process is dynamic, with the blue light area slowly advancing, repelling our "couriers" to carry the virus to our desired area. The effluent will then enter the detecting area for virus concentration testing.
(Figure 4. upper sampling zone, containing pre-treatment chamber, enrichment chamber)
On the middle level, we place six sample ports, which hold some of the " detector " solution. These ports are replaceable, and after one sample is taken, the entire chamber is rotated 60° for the next sample to be taken. Procedures are in place to ensure adequate reaction time.
After a period of time, the fluorescence detection device is automatically turned on to detect fluorescence intensity and automatically anticipate reports of the number of infections in the service area based on the modeling results. The results are reported directly to the epidemic prevention and detection department.
After a round of testing is completed, the testing waste solution will be centralized and disposed of by specialized personnel.
