Contribution
Experiment
- 1. We provide a detailed protocol for IPTG induced protein expression and affinity chromatography purification
- 2. We provide a complete experimental procedure for genetic engineering of Bacillus subtilis, including bacterial culture, plasmid transformation, and protein induction, and offer a method for gene knockout of Bacillus subtilis using the knockout plasmid pJOE8999.
- We provide a new method for inducing biofilm formation in Bacillus subtilis and propose methods for detecting bacterial adhesion and biofilm adhesion to materials
Parts
- 1. We have extended and applied the surface display system of Bacillus subtilis, proposing a whole cell immobilization platform to promote the application of synthetic biology in wastewater treatment.
- 2. We propose a novel suicide switch circuit that uses density as the control switch, which has a wide range of applications in biosafety and provides a new design concept for suicide switches for other teams.
- 3. 26 Composite Parts and 17 new Basic Parts were uploaded.
Modeling
- 1. Strategic Optimization: We mathematically simulated and identified the superior bacterial adhesion strategy (SpyTag-SpyCatcher system), demonstrating its advantages in biofilm stability and metabolic efficiency.
- 2. Quantitative Biosafety: We developed a multi-model (ODE/PDE/fluid cloak) simulation framework to validate the density-dependent suicide switch, providing quantitative thresholds and robust containment strategies for synthetic biology applications.
- 3. Efficient Protein Design: We applied a lightweight, score-based protein modeling approach to rapidly rank TasA fusion constructs, predictively guiding the design of variants with enhanced polystyrene adsorption capacity.
Human Practices
- 1. We built a demand-driven background research system by integrating data from the UN World Water Report, Sichuan's "14th Five-Year Plan" for heavy metal prevention, and Chengdu;s wastewater treatment infrastructure. It clarified the core bottleneck of "microbial loss" in engineered bacteria industrialization and focused the project on Sichuan's prominent cadmium pollution, laying a practical foundation for the TasAnchor platform.
- 2. We established a multi-dimensional stakeholder collaboration mechanism covering five groups. For the public, we designed a hierarchical science communication system. For wastewater treatment enterprises, we adjusted the application positioning to "tertiary advanced treatment of high-difficulty industrial wastewater" and planned comparative experiments. For the government, we added long-term ecological impact assessments to align with 2025 water ecology policies.
- 3. We realized a closed-loop iterative improvement of the project. Through three academic exchanges, we changed the chassis from E. coli to Bacillus subtilis, simplified the technical route to focus on heavy metal "detection-enrichment-elution", and prioritized the TasA-mussel foot protein fusion strategy.
- 4. We promoted inclusive science education and preserved academic heritage. We organized Sichuan University's Microbial Application and Design Competition (ACMA), launched a Braille children's science book project for the visually impaired, participated in the CCiC conference, shared experiences with iGEM teams like YNNU-China, and co-authored two manuals on synthetic biology that are easy to share.