Team:NCTU Formosa/Collaborations
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NTHU collaboration
Figure 1: We held experiments together at both NCTU and NTHU
Early July, we appointed a meet up with NTHU, a team that has long been our friend. We then shared our project and discussed how we could collaborate upon knowing that NTHU Taiwan was focusing on agriculture this year. We quickly came up with the idea to help them reach out to iGEM team NCTU Formosa 2018, our senior iGEMer, who also developed an intelligence agriculture-based project. Besides, NTHU Taiwan asked us to provide our modified E. coli, which contained ydfD toxin to test toxin gene-based biosafety design.
Moreover, we requested them to feedback their toxin gene test data as a supplement to our experiment results. The result confirmed our concept for which the toxin gene could not entirely kill bacteria. Furthermore, they brought us a membrane they developed that could filter bacteria, and we tried to help them test its function.
NCTU to NTHU
We have assisted them in reaching out to intelligence agriculture experts. Moreover, provide our modified E. coli contained ydfD toxin for them to test toxin gene-based biosafety design. Also, help them test their special designed membrane.
NTHU to NCTU
They helped us confirm our concept, which toxin gene could not entirely kill bacteria, and provide their data as a supplement for our experiment result.
Result
Figure 2: ydfD functional test from NTHU
Base on the result(Figure 1), we could observe that the growth curve of bacteria was slightly slow down after 100 minutes, which indicated that the toxin gene had suppressed the growth of bacteria after it expressed, but their result was not as similar as we did. In our experiment, the final O.D. value reaches two after 7 hours of detection, but O.D. value they detected only increases to O.D. 0.7 after 7 hours of observation, which may lead to our concern that the bacteria did not grow as well. We inferred this difference might occur due to different measuring conditions.
Mingdao collaboration
Mingdao to NCTU
This year, we had collaboration with Mingdao high school by giving them some suggestions to their presentation and helped them do SDS PAGE to confirm that their target protein could express. On the other hand, they did a functional test for us to confirm that our toxin ydfD could work usually.
Result
Figure 3: ydfD functional test from Mingdao
Clones 1 & 2(Figure 2) grew smoothly as usual without IPTG induction and were set as control groups. Clones 1 & 2 with IPTG expressed the toxin of ydfD and grew a little slower than the controls and the O.D. value difference is not as much as our experiment data. However, the growth speed instantly increased around the OD of 0.7 at the time of 250 minutes. The result suggested the toxin affects the growth of E. coli and might make an adaptive mutation in E. coli in the pressure of the toxin.
NCTU to Mingdao
We conducted SDS-PAGE with the Bacillus total lysates prepared by us. The gel stained with coomassie blue was presented below. Lanes 2 & 3 showed CYP2E1 with different cofactors (5-aminolevulinic acid and 5-aminolevulinic acid plus FeSO4, respectively) at ~55 kDa. Due to many other proteins were similar in size (see Lane 1 for the WT control), although with a thinner band on the gel, the CYP2E1 expression and the protein induction conditions needed to be further examined.
Figure 4: SDS page result
Most Asked Questions About Genetic Engineering
We were invited by Queen's University Canada for this massive collaboration, which involves seven iGEM teams around the world. These seven teams used modern technology combined with their knowledge trying to answer the questions that people might have about genetic engineering. Thus, a Q&A video about the most asked questions about gene engineering was shot and bought to the internet, free for everyone. The aim of using new media was to achieve user-friendly and gain most acceptance with general students sharing academic information, in the kindest way. We understand the importance of public acceptance of genetic engineering; that's why we acted!.
We also have other collaborations in teaching machine learning techniques to Tokyo Tech. Furthermore, US_AFRL_CarrollHS consulted us with Wiki page design, and we responce them by E-maIl.
Figure 5: The seven teams that making the video.
