1. Point of view: using part BBa_K592100 Blue Fluorescent Protein (mTagBFP) as report gene in cyanobacteria
This year, we decided to choose the mTagBFP BFP(BBa_K592100) as report gene to test promoter in cyanobacteria.
In our project, we sought to identify the most suitable reporter gene for use in cyanobacteria, particularly for promoter testing. Cyanobacteria are unique among prokaryotes because of their extensive photosynthetic apparatus, which contains pigments such as chlorophyll and phycobiliproteins. These pigments naturally emit strong fluorescence in the green and red regions of the spectrum. As a result, when commonly used reporter genes such as Enhanced Green Fluorescent Protein (EGFP) or Red Fluorescent Protein (RFP) are expressed in cyanobacteria, it becomes challenging to distinguish the reporter’s signal from the background autofluorescence generated by the host cells themselves.
To address this challenge, we decided to use Blue Fluorescent Protein (BFP), specifically mTagBFP, as our reporter gene for promoter activity assays in cyanobacteria. The key advantage of BFP lies in its unique excitation and emission properties. mTagBFP is excited at around 380 nm and emits fluorescence at around 440 nm. This spectral profile is well separated from the intrinsic fluorescence peaks of cyanobacterial pigments, which are concentrated primarily in the green and red wavelengths.
Excitation and Emission of Fluorescent Proteins
https://www.xepu.us/about/excitation-emission-fluorescent-protein.html
2. Application of carbon nano dots on Moss Physcomitrella patens
The carbon nanodots transformation system provide an de-novel way of plant genetic transformation, on the past 6 years, we continued working on it and gain numerous progress.
| Year | Contributions in plant Synbio |
|---|---|
| 2020 | Biopesticide targeting locust: SZ-SHD 2020 |
| 2021 | Update works about 2020 project and add information to part BBa_K3686010 about its' application in carbon dot transformation: Part:BBa_K3686010 |
| 2022 | Discover the application and design experiment protocol for outer membrane vesicle(OMV) protein: SZ-SHD2022 Engineeringthis contribution has been used by Imperial-College team this year. |
| 2023 | intelli-P plant sensor project: SZ-SHD2023 Engineering |
| 2024 | TTTT system; new model plant; enginnnered cyanobacterica SZ-SHD2024 plant |
Five years of SZ-SHD in plant synbio
SZ-SHD2020 project: insecticidal missile
SZ-SHD 2020
a.Overview of 2023 SZ-SHD phosphate phytosensor(2023 best plant synthetic biology project); b. low noise amplifier(BBa_K4844005, 2023 best composite part); c. live visualization of eyGFP(UV) under UV flashlight To test the real-world application of our product For more information, visit: SZ-SHD2023 engineering
This year, we using carbon dots meditate transient transformation on moss to express two functional protein: AtPCS and PseMTs to chelate and reduce the heavy metal ion in water to metal particles.
Establishment of sterile Physcomitrella patens cultures from non-sterile laboratory stock. Moss was transferred to BCD medium containing 100 mg/L timentin and incubated at 25°C with 5000 LUX continuous light. Left: Moss fragments at Day 0. Right: Proliferation of sterile moss after 14 days.
Moss expressing vector
Environmental scanning electron microscope (ESEM) images of Physcomitrella patens tissue after metal nanoparticle biosynthesis. Moss samples were fixed with 2.5% glutaraldehyde and imaged using a Quattro ESEM at 5.00 kV accelerating voltage, 57 pA beam current, 5.4 mm working distance, and secondary electron detection (SE mode). Nanoparticle aggregates (red arrows) are visible on the moss surface at 1,000× magnification (left, scale bar: 40 μm) and 2,500× magnification (right, scale bar: 10 μm), confirming successful metal nanoparticle formation.
More information on: engineering
3. Design and Development of Our Bioreactor Hardware
In designing our bioreactor hardware, we were inspired by the continuous directed evolution system developed by the our team in 2024 ( hardware).Our system effectively demonstrated how bacteria could be forced to adapt to environmental stressors through a continuous culture device, using peristaltic pumps, magnetic stirring, and multi-chambered bottles to achieve directional evolution. Building upon this foundation, our team set out to engineer a bioreactor specifically optimized for cultivating cyanobacteria with greater efficiency, adaptability, and ease of use.
For more information and 3D printing file, visit: hardware
4. Full open access of research material guarantee
We deeply recognize the importance of open sharing in scientific research. To support transparency and collaboration, all of our plasmid files are freely accessible not only through the iGEM part registry, but also on our dedicated protocol page:
protocol
In addition, we are pleased to offer our experimental materials—including the sterile moss Physcomitrella patens strain, cyanobacteria PCC6803, and our plasmid constructs—to the iGEM community and other researchers upon request. We are committed to supporting scientific progress and welcome any inquiries or requests for materials.
Please feel free to contact us for any further needs or collaborations.