1. Adding documentation of promoter CAMV 35S (BBa_K1467101)
This year our team aims to develop a "phytosensor" that revolutionizes the way we monitor phosphate levels within plants. ( Design ) To achieve this, we integrated our patented carbon nanodot-based tracked, transformation, translation, and trans regulation(TTTT) technique ( Plant synthetic biology ) and a multi-level gene circuit designed by our team.
Therefore, the CAMV 35S plant promoter was used throughout our whole project. There are some key introductions to our application.
CAMV 35S as a promoter for our next-generation Plant report gene eyGFP_UV and used as a positive control
The traditional equipment to detect GFP is not only highly expensive but also requires lots of training for operation. Thus, we are excited to introduce a new plant report gene, called eYGFP-uv (a GFP variant, BBa_K4844000), in transient expression we observed bright fluorescence under UV light on tobacco leaves. eyGFPuv was optimized for maximal fluorescence to be observed by naked eyes under UV light instead of using a fluorescence microscope.
Our transient expression vector was built up with a plant-strong promoter CAMV 35S, a 5'UTR on the upstream of eygfp (uv) to enhance the translation efficiency in plants and fused with a 3*flag tag downstream for western blot detection of the protein. (Our vector sequence can be downloaded at the supplementary material page Experiments )
More information about this application can be found at: Engineering
Recombined CAMV 35S work in our amplifier gene circuit
In order to reduce the leaking expression (background expression without low phosphate pressure) of the original low phosphate promoter we used and increase the promoting strength. We decided to design a signal amplification system.
More about this design can be found at: Engineering
In our gene circuit, a recombined 35s promoter was used for the out put of our report gene.
Our qPCR result indicates that our low noise amplifier part can not only increase the expression strength but also reduce leaky expression.
Other potential applications of 35S:
Based on our carbon nanodot-based tracked, transformation, translation, and trans regulation(TTTT) system, more potential applications can be done. Learn more at: Plant synthetic biology
2. Design of intelligent phosphorus fertilizer system
Our inspiration came from the team Nanjing_high_school 2020. In our design, the system modified engineered bacteria that respond to low-phosphate stress signals emanating from plant roots. When activated, these bacteria produce and secrete gluconic acid, a naturally occurring compound renowned for its ability to enhance the solubility of calcium salts in the soil. This ingenious mechanism effectively mobilizes otherwise insoluble phosphate salts, rendering them accessible to plants in a form they can readily absorb. By dramatically enhancing phosphate utilization in the soil, this system serves as a game-changer, not only significantly boosting crop productivity but also mitigating the environmental repercussions associated with excessive fertilizer application.
