Background:
During our iGEM research we realized that recruitment of Syk resulted in a several fold reduction in signal rather than an increase in signal. This is problematic because the signal is already weak and we would like to avoid having a logic inverter. We suspect that this change is due to the long 15 aa Syk-TEV linker and no additional linkers around the TCS. In this experiment we are optimizing CD79A and CD79B linkers for the 15 aa Syk or Lyn cofactor. We plan on making an X shaped pattern on an N terminal linker length vs. C terminal linker length plot and using those data points to extrapolate the ideal linker lengths at which point we will do a more precise screening.
Parts Needed:
- h:Gmab H
- h:Gmab L
- h:CD79A
- h:CD79B
- h:CD79A-TCS-Gal4VP16
- h:CD79B-TCS-Gal4VP16
- T/h:Lyn/Syk-TEV????
- h:rtTA
- h:Gal4VP16
- UAS:mKate
- h:eBFP
- DOX
- aIgM
...
Procedure:
Results:
Discussion:
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