Why We Chose Kudzu As Our Project
With limited resources and equipment in the lab, our team wanted to do what we can to maximize our ability to help the community and local people. So we focused our project on the environmental issues in areas nearby. After doing some research, we learned that the invasion of plants of foreign species, specifically kudzu, has become a severe problem in the United States. This will lead to problems that will affect the agriculture of native plants, damage the electronic lines, invade private lands, and so on. These problems may cause serious consequences if people do not pay enough attention to the unlimited growth of kudzu. As a result, we came up with a synthetic biology way to control kudzu.
Designing Our Lab
After reading some papers about kudzu, we found out that there is a certain kind of plant toxin called phaseolotoxin that can control kudzu (Chen, Li, Deng, & Zhao, 2015).and, at the same time, can have a relatively small effect on the animals living nearby. Since E. coli is the only bacteria that we will be able to utilize in our lab, we decided to clone the phaseolotoxin and make the toxin expressed in the E. coli with the help of the OTCase pathway (Arvizu-Gómez, Hernández-Morales, Pastor-Palacios, Brieba, & Álvarez-Morales, 2011).
However, due to the coronavirus situation, we were not able to come into our lab for the web lab work, so we mainly focused on designing the dry lab. We are working on three models. The first model is about optimizing metabolic pathways of E. coli to avoid cell death. The second model is to simulate the spread of infectious plant diseases like Asian Soybean Rust using kudzu as a vector. In the third model, we predict how the kudzu spread and what will be the cost to gain a better understanding of the possible economic influences if we apply our project into commercial production.
Possible Wet Lab Work In The Future
In order to really produce the phaseolotoxin in our future wet lab work, we are going to use the E.coli expression system. The following is the procedure needed in the experiment.
- Sequence design (including primer design)
- PCR amplification
- Restriction enzyme digestion
- Sequencing (sending to other companies)
- Transfection (if the sequencing result is correct)
- Purification (depending on the condition of the E.coli)
Arvizu-Gómez, J. L., Hernández-Morales, A., Pastor-Palacios, G., Brieba, L. G., & Álvarez-Morales, A. (2011). Integration Host Factor (IHF) binds to the promoter region of the phtD operon involved in phaseolotoxin synthesis in P. syringae pv. phaseolicola NPS3121. BMC microbiology, 11, 90. https://doi.org/10.1186/1471-2180-11-90
Chen, L., Li, P., Deng, Z., & Zhao, C. (2015). Ornithine Transcarbamylase ArgK Plays a Dual role for the Self-defense of Phaseolotoxin Producing Pseudomonas syringae pv. phaseolicola. Scientific reports, 5, 12892. https://doi.org/10.1038/srep12892