Parts
As part of our commitment to advancing synthetic biology, we have submitted our optimized coding sequence for the barnacle Cp19k protein to the iGEM Parts Registry.
Part ID: BBa_25FRPPH9
Sequence Optimization for E. coli Expression
The original coding sequence for the Cp19k protein was derived from barnacle DNA. To enhance expression in Escherichia coli (E. coli) the modified sequence (Xu Zhenzhen, 2020):
Codon Optimization: adjusted codon usage to match E. coli's preferred codon usage, improving translation efficiency.
GC Content Adjustment: optimized the GC content to stabilize the mRNA structure, facilitating better transcription and translation processes.
Elimination of Secondary Structures: modified regions prone to forming secondary structures that could impede ribosome binding and translation initiation.
The figure below illustrates these modifications:
Top Line: Original coding sequence
Bottom Line: Optimized coding sequence
Red Bases: Nucleotides altered during optimization
These changes were designed to enhance the efficiency of protein expression in E. coli, facilitating subsequent functional analyses.
References
Xu, Zhenzhen. (2020) Expression and Stickiness Detection of Barnacle Cp19k Protein. Master's dissertation, College of Pharmacy, Hebei University.
Building a Biotechnology Lab from Scratch
Faced with the challenge of not being able to access external help, our team decided to construct a fully functioning biotechnology lab at our school, from scratch. This allowed us to complete the iGEM project independently, overcoming the limitations of unavailable resources. We sourced equipment, designed the lab space, and set up essential tools, ensuring the team could perform all necessary experiments internally and safely. This shows future generations of high school teams the possibility of completing the project independently, a notion we hope becomes more widespread in the iGEM community as we found independent, unguided exploration to be extremely rewarding.
Development of Protocols
Through the engineering cycle, we compiled, refined, and optimized a comprehensive set of protocols for each stage of the project. This process required us to critically evaluate existing methodologies, adapt them to our specific needs, and share the final protocols with the iGEM community for future teams to benefit from.
Fostering a Biotechnology Pathway
Recognizing the importance of practical experience, we launched a Biotechnology pathway at our school, aimed at engaging students from Year 9 to 13. Starting with the Bronze Crest Award and Biotechnology CCA and culminating in participation in the iGEM competition, this pathway is designed to nurture young talent in biotechnology and bioengineering research. Giving students the opportunity to explore hands-on science, hypothesising and dicussing results, we intend to inspire future generations and cultivate a passion for these fields from an early age.