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Team:Brown/Notebook Meetings/6-8-09
From 2009.igem.org
iGEM Meeting
June 8, 2009
- 6:00 PM PST
- 6:19: We discovered we had video chat capabilities. It made the meeting AWESOME.
- 6:23: Possible 3rd kill switch: SulA stops FtsZ gene which is responsible for forming a contractile ring in binary fission
- Another possibility: having a cell synthesize a colicin under certain conditions. Colicin is a toxin that kills bacteria. Check out this Pubmed article. **http://www.ncbi.nlm.nih.gov/pubmed/17347522
- 6:28: Will discusses the EV131 failed clinical trials. The Histamine Binding Protein needs to not be in the bloodstream because it works outside of the bloodstream to cause vasodilation. and the IGE Antibody binding protein needs to be in the bloodstream to bind. This defeats our bifunctional protein idea.
- Will’s new tactic: we could create a safe strain of S. aureus that secretes EV131 in the nose, and focus on establishing a secretion protocol. We can make biobricks for secretion. We can create new parts for the bacteria. Will is making a list of other things we can focus on. It’s on the google group.
- Something like this:
- Going from:
- Cambridge's project to make a gram-positive chassis: https://2007.igem.org/Cambridge/Gram-positive_chassis_project
- The Parts page about B. subtilis: http://partsregistry.org/Chassis/B.subtilis_Strains
- Use Bacillus subtilis as our chassis.
- Figure out how to turn off the genes that allow for mobility.
- Add genes from S. aureus that allow B. subtilis to cling to mucus.
- Add new parts for protein secretion in gram-positive bacteria (in addition to the one already there)
- Come up with a new technical standard regarding gram-positive bacteria as a factory for proteins (this hasn't been done, I think)
- Biobricks for uptake into blood
- Indu will email professor Wessel about the bifunctional issue. We will go from there.
- New issue, the bifunctional protein is that we are targeting to very different things, with different properties, quantities, and location. The IGE Antibody binding protein may be too large compared to the histamine binding protein, and this protein may be too large to be taken into the bloodstream. This can be solved by finding the single important region of each receptor and bridging them together. Protocols and biobricks for secretion, production, protein bridge can be our focus.
- 7:02: We need faculty input on these issues! Michael will follow up with Wessel and Adrian about the June 12 meeting.
- 7:08: Indu and Steph are researching parts of the IGE Antibody that we can focus on to minimize the size of the binding protein
- 7:20: We need to order / obtain UCP, Holin, colicin, and SulA. There are many different holins to choose from; Ahmad wants us to pick a couple! Ahmad: randomly order a couple from the Texas A&M professor, just so we have them on hand. Get the sequences.
- We can characterize the one Holin in the registry and see if it works for our project. We can do the same for UCP. SulA Promoter (?) and colicin are in the registry…
- 7:25: For the first meeting at 8:30 AM on Monday, we will be talking about project plans and splitting up into teams.
- 7:38: We still need a repressor for the entire Safe Cell system!! We should consult Wessel about repressors. Gotta be tight, not endogenous to the organism, and uncommon in the world. Michael will send him an email.
- 7:43: Time and money about protocols are a concern. We can ask Adrian to take a look at the protocols and advise us step by step.
- 7:47: Everyone send out your emails! And forward the replies to the entire group. See you Monday at 9 AM Eastern Time.
