JUDGING CRITERIA

In fulfillment of the requirements for the Gold Medal, the 2016 Wageningen iGEM Team did the following:

Bronze Medal Requirements：

1. Completed the registration requirements and Project Summary form. Prepared and will present a poster and talk at the 2016 Jamboree.
2. Entered all necessary information detailing 39 BioBricks into the Registry of Standard Parts.
3. Designed parts in conformity with accepted BioBrick standards.
4. DNA for 39 BioBricks entered in the Registry has been sent in to iGEM headquarters. The majority of these parts were confirmed with DNA sequencing.

Silver Medal Requirements：

1. Demonstrated that several submitted new biobricks work as expected:
• The composite of chitinase A and B (BBa_K1913002, BBa_K1913003) from S. marcecens. These chitinases have been demonstrated to have relative specific toxicity to V. destructor. we modified these chitinases gene and made them into biobricks.
• The composites of Vitamin B12 and Guanine riboswitches (BBa_K1913008, BBa_K1913009) from E.coli K12 and Bacillus subtilis respectively, which negatively modulate translation upon vitamin B12 and guanine binding. These composites with RFP reporter, was shown to be functional in vivo.
• The composite of cry3Aa toxin (BBa_K1913015) from Bacillus thuringiensis Bt886, this gene encodes an 85.78-kDa, cry3Aa1 like protein that has a high level of toxicity towards most kind of insects. We tested its toxicity in vitro by using brush border membrane vesicles (BBMV).
FixK2/plac hybrid promoters (BBa_K1913023, BBa_K1913022) are integrated part of natural FixK2 promoter sequence from B. japonicum with common FixJ binding site and lac operator. It could be either activated by YF1-FixJ generator (BBa_K1913034) or inhibit by lacI.
2. Considered safety in our project by implementing a light Kill-Switch system and a synthetic amino acid system to prevent horizontal gene transfer. By implementing these systems, we will reduce the environmental impact of BT.
3. Collaborated with Groningen university, Delft university iGEM team to test and characterize new biobricks from each other. Delft iGEM team assisted us to make electron microscopy image of membrane vesicles, we helped them to characterized different fluorescent protein. Also, we helped Groningen iGEM team to sequence their biobricks and they tested our riboswitch device in return.
4. For the human practice part, collaborated with Synenergene and discussed our project application scenario and a techno-moral vignette so that we created a magazine about the future techno moral consequences and social impact on our product. In addition, contacted various Dutch newspaper and magazines to publicize our project and synthetic biology widely and participated in several conferences to inform different stakeholders about our project and collect feedback on our project application. Also, organized the national conference with Dutch iGEM teams to communicate and share own iGEM experience.

Gold Medal Requirements:

1. Improve and verify several previous parts:
• Improved upon the existing set of quorum sensing composite part (BBa_K546005) by reducing an additional luxR coding sequence and made a better characterization results of this system.
• Designed and provided new FixK2 promoters by adding core control element and typical FixJ binding sites and operons, making them could be controlled by different inputs signal. According to our experimental results, the original FixK2 promoter (BBa_K592006) in iGEM part registry has lower intensity of transcription compared to our new hybrid FixK2 promoter. Thus, we created a new functional FixK2 promoter.
• Verified the existing set of secretion system composite (BBa_K215108) by re-sequencing the full sequence.
2. Integrated Human Practices:

We consulted and communicated with several stakeholders and specialist such as beekeepers from the Dutch beekeeper association, specialists from the university to obtain their opinions and suggestions in order to make our BT product to be more feasible and acceptable. According to their suggestions of Dutch Beekeeper association, our product should fit with hobbyist character of beekeeping and never affect honey production. Therefore, we amended our project design in the following two aspects. First of all, the existing insecticide-acaricidei are prone to result in resistance of mite if it could not be applied in an appropriate way. In order to prevent development of resistance to our specific anti-mite toxin, we used two kinds of riboswitch and quorum sensing system to make sure our toxin production keep within a reasonable range. In addition, we collaborated with Design Academy Eindhoven to design a tea bag like product that beekeeper can apply it before the beginning of winter to feed bees with sugar water, which could adapt to hobbyist character of beekeeping and avoid interference of honey production.