iGEM NTHU 2024, iGEM CSMU x NCHU, iGEM CCU
We proactively reached out to other iGEM teams in Taiwan, including iGEM NTHU 2024, iGEM CSMU x NCHU and iGEM CCU, inviting them to share our projects through online or in-person meetings. We practiced presenting our project within a limited timeframe. These exchanges provided us with valuable feedback and suggestions while also deepening our understanding of each team's circumstances.
Questions about Lpp-OmpA
Our Wet Lab has had some questions during the mid-phase of our project: how can we assess the effectiveness of Lpp-OmpA in expressing heterologous proteins in the outer membrane of Escherichia coli? Does Lpp-OmpA shows any differences in expressing various heterologous proteins, and what kind of protein does Lpp-OmpA prefer to express?
To find out the answer, our team reached out to other iGEM teams whose project is related to ours. By having online meetings with other teams, we shared project, asked questions and gave suggestions to each other.
iGEM NTU Taiwan
During the meeting with NTU Taiwan, we learned that Lpp-OmpA might show varying expression levels depending on whether Lpp-OmpA is connected to the N-terminal or C-terminal of the heterologous protein. Also, they were concerned about our plan to overexpress Lpp-OmpA, worrying that the excessive transmembrane proteins could lead to the rupture of E. coli’s cell membrane. However, we conducted experiments and confirmed that our expression levels do not cause E. coli to rupture.
WHU iGEM
WHU iGEM, similar to us, is employing Lpp-OmpA to express proteins on the E. coli cell membrane. While having our online meeting with them, we raised several questions about Lpp-OmpA, and WHU iGEM generously shared their insights. They employed Western blotting and flow cytometry to confirm that heterologous proteins were accurately expressed on the membrane. Additionally, they provided information on how Lpp-OmpA compares with other membrane proteins. This exchange had a significant impact on the progress of our project.
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