Team:SZU-China/Contribution

We got the recombinant plasmid “pSC101-TALEsp2-tnaA-Fmo “containing the sequence of the composite part BBa_K3264022 from our partner Shanghai_SFLS_SPBS. And the composite part contains basic BBa_K1131000 for gene fmo. The whole composite part is supposed to express tnaA encoded tryptophane and fmo encoded Flavin-containing monooxygenase (FMO) for catalyzing tryptophan into indigo.

1.1 Experimental Characterization

We confirmed the function of the composite part in successfully expressing the tryptophanase and FMO for transforming tryptophan into indigo by labwork.

After flask-shake cultivation and centrifugation, the cell pellet was resuspended by dimethyl sulfoxide (DMSO), and tested by ultraviolet-visible spectrophotometer UV-2250 for optical density at 620 nm (Fig. 1). Comparing the sample OD620 with the indigo standard curve, wedetermined our indigo produdction as around 0.08 mg/ml.

Fig. 1 Bioidigo absorption spectrumand crystal blue solution of our redissolved indigo in DMSO

1.2 Documentary Characterization

To better help future teams understand the briliant mechanism of FMO in the pathway of bio-indigo synthesis, as well as the history of indigo, we did some literature research and document relevant information on the Registry page of BBa_K1131000 & composite BBa_K3264022.

The information include the background intro of FMO and indigo, biochemical characteristics of FMO, scheme of FMO in bio-indigo synthesis (Fig. 2), and relevant creative experiments from literature.

Fig. 2 Catalytic cycle of flavin-containing monooxygenase

We believe our documentary characterization can offer a quick overview for future teams about FMO and indigo, which can help them better locate valuable message for own interests to search for more.

2. Create a New Way for Project Performance and Collaboration

In June, 2020, we held the 4^th Southern China Regional Meeting. To keep social distancing during the global Covid-19 global pandemic, we transferred the meeting from offline to online. And for easier information exchange, we utilized the special column of Bilibili as a platform for project exhibition among different teams. We successfully built up the first community of Chinese iGEMers in this streaming entertainment service popular among young people (Fig. 3). Our successful experience can be the template for the future teams to organize online collaboration and exchange information via Bilibili.

Fig. 3 Screens showing the videos submitted by iGEMers on Bilibili in our campaign

Merits of the our new way for interacting on Bilibili

Every team can set up their own accounts in Bilibili for sharing videos for free.
The user account can be inherited by the school’s following new iGEMers, and they can use it as a official account for promotion and document year by year. And the set contact way can benefits future team collaboration.
Before a meeting, related teams can share their project intros and videos on the same column for efficient mutual information exchange (Fig. 4).
Future iGEMers and the public can have easy access to the previous videos offered by former iGEMers, which is a marvelous way for propagating the idea of synthetic biology.

Team:SZU-China/Contribution

Contribution

Fig. 1 Bioidigo absorption spectrumand crystal blue solution of our redissolved indigo in DMSO

Fig. 2 Catalytic cycle of flavin-containing monooxygenase

Fig. 3 Screens showing the videos submitted by iGEMers on Bilibili in our campaign

Fig. 4 Intro and video of a team on our special column of Bilibili