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Revision as of 17:12, 18 September 2015
As the famous Chinese proverb goes,"Union is strength". It adequately shows the importance of collaboration. During the past years, team Nankai has been cooperating intimately with team Tianjin, our beloved neighbor and friend. We discussed from time to time about different details in our projects and helped each other out when it was needed. Besides, this year team Nankai set up a new resource sharing platform named"iShare". It's a platform for all the iGEM teams to get what they need. To gather more the information, we have also reach for many other teams for help. Thanks to all the iGEM teams who offered us help, ishare is now fully functional.
Collaboration
I. Collaboration with our neighbor TJU
The Lotus Festival held at TJU
In early April, when the lotus is the most exuberant in school, team Tianjin held a lotus festival to popularize the knowledge of biotechnology. As their neighbor and partner, team Nankai were invited to the festival. We thanked their hospitality and offered them several samples of actinomycetes for exhibition. During the festival we had some nice chats with members of team Tianjin and had a good time enjoying the beautiful view inside the campus of Tianjin University.
The front gate of TJU.
Poster for lotus festival.
Group photo at TJU.
Supporting date of hydrophobin expression
This year, iGEM team Tianjin worked on a special small protein called hydrophobin. They tried to optimize the expression of hydrophobin in prokaryotic expression system. However, a special member in hydrophobin group called injanus, also known as HGF1, could not be expressed in E.coli. They still needed the dates about the expression of injanus and thus reached us for help. Fortunately we managed to characterize the expression of injanus in Pichia pastoris,with the help of Qiao's laboratory in Nankai university. The results were sent to team Tianjin for further employment.
We employed fed-batch fermentation to produce injanus (HGFI), and injanus produced was detected in fermentation broth through HPLC (showed in Figure 1 and Figure 2). The spectra peak of pure injauns appeared around 30-32 mins, which showed the spectra peak appeared at around 33 mins in HPLC profiles of fermentation broth was injauns.
Figure 1. HPLC profiles of pure injanus.
Figure 2. HPLC profiles of fermentation broth.
Figure 3. Silver staining along with SDS-PAGE (a) and western blot analysis of fermentation broth (b). Injanus fermentation broth (Lane 2,3), pure injanus (Lane 1).
We then analyze the broth though SDS-PAGE and western blot (showed in Figure 3). According to the results, molecule weight of injanus was around 8kDa and the injanus fermentation broth was mainly in two states, monomer and dimer. Dimer injanus binds better with the primary antibody of injanus.
In addition, we compared the production of injanus after fermentation in bioreactor and flask cultivation. The concentration of injanus in broth was measured by UPLC, and the standard curve of concentration with adsorption area of UPLC was measured using pure injanus samples. Figure 4b shows the cell fresh weight and injanus production in different methanol induction days (Figure 4b). After 4 days’ induction, in bioreactor the cell fresh reached 365g/L and in the meantime the injanus density reached 0.3g/L, which was about 10 times the production of injanus in flask cultivation. Results of SDS-PAGE indicated that the production of injanus kept increasing during the 4 day’s induction (Figure. 4a).
Figure 4. (a) Silver staining along with SDS-PAGE of injanus fermentation broth in different induction time (Lane 1-4 corresponding 24,48,72,96 hours). (b) Injanus production and cell fresh weight in of different induction time.
Ⅱ. Collaboration with other teams for iShare
IShare is a brand new idea raised up by Nankai Team in 2014.It is an information-sharing platform aiming to converge all information of iGEM China teams involved in our experiment such as Topic, Introduction, Materials and Methods, Progression, Parts.In a word, it will be a platform which is beneficial for all the igem teams.
Therefore, during the summit of domestic igem teams held by Beijing University, we seized the opportunity to publicize our iShare platform by playing a short video and distributing questionnaires about it. Our goal was to gain their support and information as much as possible. Excitedly, this platform was supported by most present teams and we exactly got much information and advice, which give us much confidence and encouragement for building this platform!