Collaborations

The educational experiment we designed today involved using colorful fluorescent bacteria to paint panels to inspire young people to get excited about life sciences.

We need 4 different colors of fluorescent proteins, including GFP, RFP, YFP and BFP.

GFP

Since we are a new iGEM team, we do not even have a basic component library, and we are not using GFP in our project this year, so we are looking for GFP plasmids from outside.

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

RFP

Regarding the source of RFP，the iBowu-China team gave us a clue and their mentor told us that we have the corresponding plasmids from BGI and also helped us to contact the corresponding person in charge. Thanks to their generosity, they gave us mCherryFP directly instead of RFP. Here the process of validation and transformation was saved for us, and we just proceeded to shake the bacteria directly.

BFP

In the process of searching for the final BFP, we found that SHSBNU-China was also searching for the BFP, and we learned from the clues they gave that BIT-China had used the BFP in previous years, so we met the captain of BIT-China through the WeChat group in China, and the captain of BIT-China kindly sent us a plasmid It is a plasmid that contains the mTagBFP plasmid.

Looking at the DNA profile of this plasmid, we found that there was no promoter on this plasmid, so we designed our own primers to obtain the mTagBFP part of the gene and then transferred it to the pet28a plasmid that we used in this year's experiment. 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. But we will not be able to catch up with our educational experiment activity of painting.

Summary

For the final educational activity, we are using both GPF and mCherryFP plasmids for transformation and plate coating.