Team:XMU-China/Collaborations
No one can make great progress without advice and golden ideas from their excellent partners. During last six month, we participated in some iGEM conferences or even worked as the innovative online meeting organizers. Hopefully these events contributed greatly to all of us.
From August 19th to 23rd, our team XMU-China participated in the 6th CCiC (Conference of China iGEMer Community) conference held by Shenzhen Advanced Technology Research Institute of Chinese Academy of Sciences. The theme of the conference was “Automation & Innovation” this year.
During the conference, through poster display and social communications, we actively communicated with other teams and gained a lot of help from them. Fortunately, we have established cooperative relations with HUBU-WUHAN, NEU-China and UESTC-China. In addition, since we tried to use the CAV1 gene derived from human in improvement part, we consulted SEU-Nanjing-China about the common problems and solutions of expressing heterologous genes in Escherichia coli.
We received many suggestions from the judges after exhibition the design and advance of our project during CCiC. Therefore, the design of our project were improved after the conference. In the end, the story of “Gone with the Wind” was selected as a carrier, and the engineering bacteria was used to demonstrate group relations.
The organizers also set many lectures to deepen our understanding of the development of synthetic biology. The two teams (GreatBay_China and SZU_China), who won the iGEM Final List in 2018, were selected to give presentation. Their experience in team management and competition give us positive mindset and more confidence in face of our future developments and final Jamboree.
Some of our team members participated in the iGEM Southern China Regional Meeting which was held at Shenzhen University this year's. The meeting attracted more than a dozen teams from south China to introduced their project design and prompted us to establish contact with some other teams after the meeting.
An idea come from brainstorming stage of our team, which use alkaloids to kill fouling organism barnacles, was surprisingly similar to the design introduced by Greatbay_SCIE in the meeting presentation. We had more in-depth communication with Greatbay_SCIE team in design of the genetic circuit. As we hope to construct innovative project, the conmunication encourage us to explore in a new field through brainstorming. The meeting was a great impetus for us to turn our attention to the study of sociology using synthetic biology.
DUT_China_A iGEM
It is meaningful and time-saving to guide us in operating our experiment through establishing an appropriate model. This year, team DUT China A contacted us to exchange our similar modeling ideas in 2018 XMU China. We shared our ideas when we built the model in last year, and we gave some guidance which help them successfully predict their sequence. It is supposed to be beneficial to the development and improvement of the project in the long run.
HUBU-WUHAN iGEM
This year, HUBU-WUHAN team aims to build up biological parts in Zymomonas mobilis to convert waste cartons into Poly-β-hydroxybutyrate (PHB) and biofuels. The cellulosome was expressed in the Z. mobilis to construct CBP strains that can utilize cellulose directly, because cellulose cannot be used as a single carbon source in medium. It’s similar to “Cooperative” module in the design of XMU-China team: let different groups of E. coli degrade cellulose for living, but the cellulases and experimental methods we used were different. After watching their project during the 6th Conference of China iGEMer Community (CCiC), we communicated each other about the issues related to cellulases.
After CCiC, we met some trouble when determine enzymatic activity of our exoglucanase (Cex), though the protein were proved to be expressed successfully by SDS-PAGE. However, we successfully solved this problem by using 4-Methylumbelliferyl β-D-Cellobioside (MUC) few days later. Meanwhile, HUBU-WUHAN team also faced problems in the determination of their exoglucanases (CBH I and Cc Exgs), and then asked us for some help.
Thus, we provided our engineered E. coli with cex gene to them as a positive control to help them solve problems in their enzyme activity determination process. E. coli with bgl1A gene coding beta-glucosidase were also sent to them as a supplemental part since they did not have enzyme to degrade cellobiose, one of the intermediate products in cellulose degradation. They also mailed their plasmids with gene coding CBH I and Cc Exgs to us that we can determine enzymatic activities of CBH I and Cc Exgs in our method.
We transformed their plasmid, which contains gene related to spectinomycin resistance, into E. coli BL21 (DE3) and selected suitable single colony for large-scale culturing by using spectinomycin they provided as inducer.
Tetracycline was added into culture mediums (final concentration: 0.0 to 0.8 μg/mL, with gradient of 0.2 μg/mL) to inducting protein expression as protocol they provided. However, none of the E. coli strains with the plasmid them provided could grow up except the control (without tetracycline) (Fig. 1). Then our induction method was also used, unfortunately, negative result was obtained that we failed to induce the expression of these two exoglucanases.
Fig. 1. The growth curves of E. coli BL21 (DE3) with CBH I and Cc Exgs induced with different concentration of tetracycline (final concentration: 0.0 to 0.8 μg/mL, with gradient of 0.2 μg/mL). (A) The growth curve of the groups of CBH I. (B) The growth curve of the groups of Cc Exgs. The number of 0, 2, 4, 6 and 8 stand for different concentration of tetracycline in different groups.
UESTC-China iGEM
In present work, we have some difficulties in characterization when use two biobricks (BBa_K118022(Cex) and BBa_K118023(CenA) ) from UESTC-China to build our genetic circuits. Fortunately, we communicated with UESTC-China team about the characterization methods of BBa_K118022 and BBa_K118023So when participating in CCiC. And then we got the reference of the enzyme activity determination methods of Cex and CenA from them. At last, CenA enzyme activity was determined by using their protocol of Congo red test, and the Cex enzyme activity determination was carried out through MUC enzyme activity test based on our improved protocol.
More details about UESTC’s work can be found in https://2018.igem.org/Team:UESTC-China and https://2019.igem.org/Team:UESTC-China.
FAFU-CHINA iGEM
We communicated with FAFU-China about the protein model. They were trying to prove the two protein has a very similar function and analyze the protein structure of FLO1, but they meet some difficults in modeling and analyzing protein structure，so that we helped them building up the protein structure prediction model. We also shared some bioinformatics relevant reference with them and gave some suggestions about modeling to them. We hope that these suggestions will be useful in improving their protein model.