Overview

Our Collaborations this year consisted of plasmid resources obtained from the BNDS-China team and the BIT-China team to realize our fluorescent proteobacteria painting activity, and our participation in The Synthetic Biologic Fair hosted by KEYSTONE.

BNDS-China：GFP

Because we are in our first year of participation, we do not have the parts package that teams received in previous years, so we do not even have the basic GFP plasmid. This was the first problem we encountered.

Finally we looked for the empty vector plasmid of GFP where in BNDS-China, sfGFP-pSB1C3,Thanks to their generosity.

We performed transformation and expression after receiving the plasmids. The strain used was BL21 receptor E. coli, and after coating the plate and overnight culture, we successfully obtained the strain expressing green fluorescent protein. We sent back this result and informed them about our expression conditions.

BIT-China:BFP

In the process of searching for other fluorescent protein plasmids, we found that BIT-China had a similar goal as ours, and they also wanted to draw fluorescent proteins in their educational activities, so we contacted BIT-China through meetup's WeChat group, made our request, and obtained the BFP plasmid, which is a plasmid with mTagBFP.

However, after receiving the plasmid, we found beyond the plasmid mapping that the plasmid does not carry a promoter and cannot be expressed directly. The solution we came up with was to use what we learned from this year's experiments to design primers to link this mTagBFP section to the pet28a plasmid used in our experiments, and then use the existing experimental conditions to induce expression.fluorescence part can be expressed in BL21 with the induction material IPTG.

After completing the plasmid reconstitution experiment, we transformed BL21 with the new plasmid, T7-BFP-pet28, and found that the colonies grew very little after coating the plates. After extended incubation, the following LB plate was obtained. After that, a single colony was selected and induced with IPTG after incubation in LB medium and excitation at 399 nm and measurement of emission light at 454 nm showed that the result did not reveal fluorescence. We think it may be that the plasmid reconstitution failed.

So unfortunately, we didn't get a usable BFP in the end, and we are going to continue to optimize the original plasmid and send it for sequencing at a later stage to determine whether our construction is successful. We also gave feedback to BIT-China about this uncharacterizable result。We learned that BIT-China also gave up the attempt of this plasmid after receiving it and found that there was no promoter on the plasmid.

In the final educational activity, we used the obtained plasmids for transformation and flat painting. Thanks again to the BNDS-China and BIT-China teams. If other teams want to try the fluorescent bacteria for painting experiment, we can provide the suitable plasmids and bacterial solution for everyone.

KEYSTONE：The Synthetic Biologic Fair

We learned in the meetup weibo group that Keystone will hold The Synthetic Biologic Fair in mid-October. At this event, we exchanged project content and completion progress with several teams, including BJU-China, KEYSTONE, BHSF, and RDFZ-China.

Among them, BJU-China's art design and KEYSTONE's mini-games impressed us deeply and broadened our imagination about iGEM.

Because of COVID-19, iGEM has not had offline finals for 2 years in a row, and the lack of finals makes everyone lack a lot of opportunities to exchange projects and ideas face to face. The KEYSTONE event was just before the wiki freeze, and I observed that some university teams like UCAS even made posters, which made up for the fact that we couldn't participate in the offline finals.