IGEM:MIT/2005/End of Week Meeting: Friday, July 1
Notes from End of Week July 1
Division of Labor
- Lab Plan: Jenny, Jen
- Antibody: Maxine, Ray, Will
- Pathway: Jessica, Annie
- Let's start ordering.
- Send an e-mail of list of items we need, and then specify of location to obtain them.
- Design/spec experiments we can do right now.
- Concern of antirabbit not being expressed - Don't worry about it.
- Partial cds - did not seq. entirety. (alignment - look for obvious landmarks)
- Not using antimouse.
- We need to get the clone, we need the antibody for testing. Call them today!
- Be careful of evil people.
- Companies that make scFv's (28 wk w/ antigen, 8 wk w/o), very pricey.
- Small initial investment for large future gain.
- Manufacture DNA with fluorecein as part of sequence. Wittrup lab has done this with scFv on yeast.
- Engr own antigens - add flag to antibody, and antiflag will respond.
- Fluorecein antibody (rabbit)
- Typical protein assume = 3 nm.
- ToxR reporter and cI fusion are made. We should order them.
To talk about:
- Applications: we're thinking about it.
- ScFV antirabbit
- not expressed in cell
- delete sequence for sensing gold binding
- partial cds?
- contact author
- ScFV antimouse-too specific
- Find backup sequence?
- Call companies that manufacture ScFVs, contact authors of papers w/scfvs
- antidigoxin, antidnp; need to check out more. list of commerciall available chem that won't hurt cells?
- Santra cruz: all bind to specific aa on a human protein: problem?? check w/an advisor
- common epitopes: Myc, FLAG (found in sigma aldrich), HA (could not find)
- Paths we're going down:
- ToxR system with small molecules only
- ToxR with linker through periplasm to an OMP
- Build our own linker????
- Use the MFP (membrane fusion protein) -- use as structural support, etc.
- Use TonB
- Screen FecA linker library
- Still waiting for word from UCSF, Daniel Kuhne
- Found Sequences for these coding regions:
- ToxR cds
- ctx promoter
- alkaline phosphotase (used to test for dimerization)
- FecA, FecR
- Is attempting to use a structural connection between inner and outer membranes proteins insane?
- Maxine's antibody question