Team:TMMU China/Judging
Bronze
1. Register the team, have a great summer, and plan to have fun at the Giant Jamboree.
We have registered our team and we are prepared for the Giant Jamboree.
2. Meet all deliverables on the Requirements page (section 3).
3.Create a page on your team wiki with clear attribution of each aspect of your project. This page must clearly attribute work done by the students and distinguish it from work done by others, including host labs, advisors, instructors, sponsors, professional website designers, artists, and commercial services. Required link: https://2016.igem.org/Team:TMMU_China/Attributions
We have created a page on our team wiki with clear attribution of each aspect of our project. Work done by the students has been distinguished from the work done by others.
4.Document at least one new standard BioBrick Part or Device central to your project and submit this part to the iGEM Registry (submissions must adhere to the iGEM Registry guidelines). You may also document a new application of a BioBrick part from a previous iGEM year, adding that documentation to the part's main page.
We have documented 2 standard BioBrick Parts and submitted them to the iGEM Registry. They are BBa_K1887002 and BBa_K1887003.
Silver
1. Experimentally validate that at least one new BioBrick Part or Device of your own design and construction works as expected. Document the characterization of this part in the Main Page section of the Registry entry for that Part/Device. This working part must be different from the part you documented in Bronze medal criterion #4. Submit this part to the iGEM Parts Registry.
We have validated experimentally the BioBrick Parts BBa_K1887000 and BBa_K1887001worked as expected and they have been documented and submitted.
2. Convince the judges you have helped any registered iGEM team from a high-school, different track, another university, or institution in a significant way by, for example, mentoring a new team, characterizing a part, debugging a construct, modeling/simulating their system or helping validate a software/hardware solution to a synthetic biology problem.Required link: https://2016.igem.org/Team:TMMU_China/Collaborations
We helped the SCU-China team to test the antimicrobial activity of their designed antimicrobial peptide Cecropin XJ. We also helped the UESTC-software-China team to design some primers for their project.
3. iGEM projects involve important questions beyond the bench, for example relating to (but not limited to) ethics, sustainability, social justice, safety, security, and intellectual property rights. We refer to these activities as Human Practices in iGEM. Demonstrate how your team has identified, investigated and addressed one or more of these issues in the context of your project.
Required link: https://2016.igem.org/Team:TMMU_China/HP/Silver
In synthetic biology and iGEM, the biosafety issue is seriously considered. Another big threat to public health is the emergence of multiple antibiotic resistant bacteria. We aimed to build a markerless visual selection system for L. lactis, which would have the potential to be used as a live vector in human. However, we lacked the knowledge about the exact situation of multiple drug resistant bacteria and biological containment. Thus, we consulted Doctor Gong and Doctor Li to obtain their instruction and seek advice from them on our project design. We really appreciated the valuable advice they gave. We finally improved our project design to be antibiotic resistance gene free and biologically contained. At the CCiC conference, Doctor Zhang Haoqian gave us suggestions that we should add some quantitative results to our project, and we made adjustments accordingly. To popularize synthetic biology and iGEM, we created a special coloring book for education and public engagement.
Gold
1 .Expand on your silver medal activity by demonstrating how you have integrated the investigated issues into the design and/or execution of your project.Required link: https://2016.igem.org/Team:TMMU_China/HP/Gold
After the counseling with Doctor Gong and Doctor Li, we decided to establish the markerless visual selection system and we also chose the His segment as the targeting locus to achieve biological containment. After the CCiC, we further carried out some experiments to obtain the positive rate and the accuracy rate of the blue-white selection system. We finished these quantitative experiments and found that the accuracy rate is 100%, even only one white colony was identified in the blue colonies pool.
2. Improve the function OR characterization of a previously existing BioBrick Part or Device (created by another team, or by your own team in in a previous year of iGEM), and enter this information in the part's page on the Registry. Please see the Registry Contribution help page for help on documenting a contribution to an existing part. This part must NOT come from your team's 2016 range of part numbers.Required link: https://2016.igem.org/Team:TMMU_China/Description
The 2015 IIT Madras-India team designed the SPusp45 part ( BBa_K1654007). but no characterization data provided. Here, we provided characterization data. We found that the SPusp45 signal peptide can be used to secret protein of interest outside of the cell using the β-galactosidase as an example. The western blot result was provided. So we verified this part (BBa_K1654007) and provided documentation in the Registry. The 2015 Concrodia-Canada team designed the SPusp45 part (BBa_K1887002). After thoroughly reviewing the literature as well as wiki (the 2014 Groningen-Netherlands team and the 2015 IIT Madras-India team), we found that the SPusp45 signal peptide is of the length of 27 amino acids, but not the length of 38 amino acids as stated by the 2015 Concordia-Canada team.So we clarified this part (BBa_K1887003) and provided documentation in the Registry.
3. Demonstrate a functional proof of concept of your project. Your proof of concept must consist of a BioBrick device; a single BioBrick part cannot constitute a proof of concept.Required link: https://2016.igem.org/Team:TMMU_China/Proof
We knocked the PnisZ::lacZ::Terminator device into the His locus, and then the NZ-Blue strain appeared as blue on the X-gal plate supplemented with nisin. The NZ-Blue strain was used as the host where 5 devices with different lengths could be knocked in, and we found all the white colonies were the right ones. All the 58 white colonies derived from the mCherry device knocked in strains emitted red fluorescence after simulation by green light.
4. Show your project working under real-world conditions. To achieve this criterion, you should demonstrate your whole system, or a functional proof of concept working under simulated conditions in the lab.Required link: https://2016.igem.org/Team:TMMU_China/Demonstrate
The nisin is the inducer of the nisin controlled expression (NICE) system, but it is also an antimicrobial peptide. The NZ9000 strain can not tolerate high levels of nisin concentration. We incorporated the nisin imunity gene nisI and nisin resistance gene nsr into the NICE system, and we found that incorporation of the nsr gene into NZ9000 can make it grow at the nisin concentration of 500 IU/mL, but the NZ9000 strain can not grow at this high level of nisin concentration. We also demonstrated that the β-galactosidase protein fused with the cA domain and the SPusp45 signal peptide can be anchored to the cell wall of L. lactis.
