Team:BIT/Collaborations
Meeting up in Yanqi lake
Team UCAS hosted the meeting to communicate with other China teams，University of Chinese Academy of Sciences、Zhejiang University、Beijing Normal University、Tianjin University of Science & Technology as well as Beijing Institute of Technology attended this meeting to share and discuss the project with each other. All of the teams exchanged ideas about their project and original design, and have a quick exhibition of the wiki, poster, give some simple but classic presentation. Among which we also try to find ways to better collaborate with other teams, especially in the wet experiment、modelling and outreach. We received more than 10 useful advises about our project and having a great time!
Collaborate with UCAS
UCAS for BIT-China:
Our project’s signal transition mainly based on the GPCR pathway of Saccharomyces cerevisiae. In order to tune the system more sensitivly, we decided to optimize the promoter Pfus which is the inducible promoter of detection circuit (Pfus-mRFP-CYC1t). Team UCAS helped us to test the expression of mRFP promoted by different Pfus mutation, through the host strain fluorescence test, they helped us obtain the corresponding data. here is the link to their wiki.
BIT-China for UCAS:
In the 19th session of National Congress of the CPC, because of the limitation of the methanol(AR) procurement，the students of UCAS are in desperate need of methanol(AR) to detect their production，we help them complete their experiment by supplying them methanol(AR).
Fig.1 HPLC analysis of target compound
Fig.2 The intensity of two kinds of promoters
Collaborate with Jilin_China
BIT-China for Jilin_China:
This year we cooperated with Jilin_China, we helped them to reconfirm whether their system is stable or not.
They designed to regulate the growth of E.coli under TA system, Arabinose operon in this system controlled expression of toxin, while galactose operon can regulate the expression of the anti-toxin. According to references and pre-experiments they did, we designed the confirmatory experiments as follows:
| Plate A | Plate B | Plate C | Plate D | |
|---|---|---|---|---|
| 0.1%Ara | - | - | + | + |
| 0.1mMIPTG | - | + | - | + |
Fig.3 Growth conditions of bacteria with TA system with different inducer (after 16 hours of streak cultivation).
The results show that, arabinose can greatly suppress the growth of this strain, while IPTG shows slight suppression effects towards this strain. On the contrary, add both two inducers to the system shows no significant effect compare to wild type, indicated the expression of anti-toxin can relieve the suppression of toxin in this system.
From these experiments, we help Jilin_China to confirm their TA system can work. For further information, here is the link to their wiki.
Jilin_China for BIT-China:
Jilin_China helped us build a part, which we can test the expression of membrane protein by detect blue color(bcp).
In this process, we are concerned about whether our sweet receptor can be successfully expressed and located on the cell membrane , the strategy is to fused BCP（Blue cytochrome protein）to the N-terminal of T1R2，so that we can observe the protein T1R2 on the membrane by fluorescence confocal microscopy. The member of Jilin_China synthesize the expression cassette of Gal1 + bcp + T1R2 + CYC1t gene for us.
Fig.4 The gene of Gal1 + bcp + T1R2 + CYC1t.
Fig.5 The electrophoresis of the positive result of T1R2.
Gal1 is an inducible promoter which is induced by Galactose. bcp is blue cytochrome protein, we can observe it by fluorescence confocal microscopy. t1r2 is a membrane proteins belongs to GPCR family. CYC1t is terminator.
From these experiments, Jilin_China helped us build this part . Here is the link to their wiki.
Collaborate with Lanzhou
Lanzhou for BIT-China:
Because our project requires the expression of mRFP as signal output. We hoped to get mutations of the inducible promoter Pfus by changing the number of ste12 binding sites. Lanzhou University helped us to build a mutant Pfus . here is the link to their wiki.
Fig.6 The electrophoresis of the positive result of Pfus
BIT-China for Lanzhou:
Pectinase and celluse play an important role in their downstream circuits, they want to combine functional dsRNA with these two proteins and let them work together, which will make great sense to their subsequent appllication design.
We helped LanZhou verified the expression of their functional protein, promoting their project progress here is the link to their wiki.
Fig.7 SDS-PAGE of their target protein.
Collaborate with BIT
From July to August in the summer of 2017, BIT and BIT-China, two iGEM teams from Beijing Institute of Technology, carried on several communications about the project, daily works, the experimental processing and iGEM matters. We have discussed about problems in each other project and helped each other to solve the practical difficulties encountered in our own experiment. here is the link to their wiki.
The main content of communications between the two teams:
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