Parts:
Ig-alpha (CD79A), Ig-beta (CD79B), IgM (one of these will be fused with TEV sequence/transcription factor)
Lyn (fused with TEV protease)
Further Research:
- Lyn expression in neurons
- Disulfide bonds between Ig-alpha and Ig-beta
- Expressing parts: how big are they, can we order gBlocks?
- What antibody do we use? -> is it patented?
Preliminary Experiments:
- co-immunoprecipitation to see if Ig-alpha/Ig-beta complex is forming
- use biotinylated anti-Ig-alpha and anti-Ig-beta antibodies
- add these antibodies to "cell soup"
- run over streptavidin column
- in gel: if Ig-alpha comes off with Ig-beta, or Ig-beta came off with Ig-alpha -> suspect that they were bound
- Lyn recruitment
- fluorescence to check localization?
- biotinylated beta-amyloid to see if it sticks to cells
DNA Parts to Order:
Antibody against beta-amyloid: Gantenerumab
- light chain (220aa) -> specific to beta-amyloid
- heavy chain (~500aa) -> modified so that it is IgM rather than IgG
- Ig-alpha
- Ig-beta
- Lyn
- beta-amyloid (to test)