IGEM:MIT/2005/End of Week Meeting: Friday, July 1

Division of Labor

 * Lab Plan: Jenny, Jen
 * Antibody: Maxine, Ray, Will
 * Pathway: Jessica, Annie

Action Plan

 * 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.

Antibodies

 * 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)

Pathways

 * Typical protein assume = 3 nm.
 * ToxR reporter and cI fusion are made. We should order them.

To talk about:

Updates:
 * Applications: we're thinking about it.
 * Antibodies
 * Secondary
 * ScFV antirabbit
 * not expressed in cell
 * delete sequence for sensing gold binding
 * partial cds?
 * contact author

Questions:
 * 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?
 * Primary
 * Santra cruz: all bind to specific aa on a human protein: problem?? check w/an advisor
 * Antigens
 * 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
 * TonB
 * MalE
 * Is attempting to use a structural connection between inner and outer membranes proteins insane?
 * Maxine's antibody question