Both of team VIT_Vellore and our team focus on antibiotic resistance
issue this year, and they take efforts in improving phage therapy while we attempt to find new
inhibitors toward relative enzymes. We contacted with each other via emails and skype calls.
From the symposium held by them on Sept.4th, they interviewed expert on some questions related to our project. Dr. Ponnari Gottipati, the co-founder of Superheros against Superbugs, gave us useful suggestions on popularization of AMR. As for their comic contents, the reasons why AMR issue is serious are needed to talk with young children. It is more important to understand the concept of resistant problem, rather than the mechanism behind them. But when citizens who have already knowledge of microorganisms are the majorities, they are thirsty for knowing scientific mechanisms. As a fighter towards abuse of antibiotics, she encouraged both of us youngsters to join the alliance, no matter what kind of approach we use.
Also, it is vital for AMR fighters to be aware of serious current situation. We assisted them with spreading questionnaires among undergraduates, to collect public opinions on AMR. Since our primary enzyme is NDM-23, a mutant of NDM-1, which was found in New Delhi, India, they provided us with information on NDM-type beta-lactamases.
Although the themes of our projects differ, there are still same sections in
the process of experiment. Perhaps we use same engineering bacteria, same fusion tag in
constructions, and same experimental protocols. We didn’t fear facing difficulties, because we had
In July, when we troubled with overexpression of our interest beta-lactamases, we guessed there
about the use amount of antibiotic and the choice of competent cells. We found team Gunma
focused on E.coli
pLysS so that we communicated each other via email and did troubleshooting.
They thought our use amount of antibiotic was too much and caused death of bacteria. And they introduced the characteristic of E.coli BL21 (DE3) pLysS. Following their advice, we made some regulations then successfully expressed our beta-lactamases using E.coli BL21 (DE3) pLysS.
And after knowing their detailed project plan, we suggest that they can build relative model in order to analyze fluorescent data to reflect bacteria’s living condition.
It is brilliant to have a partner team in the same school. Although our
project themes differ, some steps are same, for example, PCR. We visited each other’s laboratory
and discussed some difficulties in the experiment, both technically and metrologically. And
since we have completed most of our experiments in September, however, they have a lot of
overlapping work to do, we assist to help them construct a gene sequence.
We have successfully completed PCR of 4 gene fragments. However, it’s a little difficult to overlap them. We have tried many times, adjusting the annual temperatures and concentration of template. Due to time limit we did not combine them finally, and this is also a valuable experience for both of us.
Gel results can be seen as right. A. PCR results of hph. Lane M, Marker, Lane1-4, hph. B. PCR results of R. Lane M, Marker, Lane1-4, R. C. PCR results of L. Lane M, Marker, Lane1-4, L. A. PCR results of RFP. Lane M, Marker, Lane1-4, RFP. E. Overlap PCR results of hph+R, succeed. Lane M, Marker, Lane1-4, hph+R. F. Overlap PCR results of L+RFP, fail. Lane M, Marker, Lane1-4, L+RFP.
We both iterate our circuits by adding GST tag to improve target protein's solubility to benefit the process of purification. And they found it difficult to purify PreScission Protease which is used to cut GST tag after affinity chromatography. So we provided our purification protocol to them since we have successfully attained protein. And finally they got this enzyme and cut GST tag from GST-AMPs.
A - Our SDS-PAGE result, showing GST-NDM-23 and
NDM-23 after being cut by PPase
B - Team WHU-China's SDS-PAGE result
From iGEM official website, we join their collaborative comic activity. We provided three photos of us (one about you running away, another about you entering a building - preferably your university with its visible name, and the last one in a building, scared). And they made their comic using our photos. Comic is an awesome way to popularize some situations, and it is innovative to combine with real photos.
They aim to discuss about the differences in the legislation regarding genetically modified
organisms (GMOs) in different countries. What is the number of varieties of genetically modified
crops approved for cultivation in your country? What is the area destined to GM crops cultivation in
And we searched for these information in our country then send to them.