Test using the whole detection circuit and Beta Amyloid to measure circuits response to input. This test will vary several circuit parameters to find the ideal sensor.
Purpose
This will determine the ideal concentration of Syk-TEVp and ideal placement of TCS-Gal4VP16 for the best signal/noise ratio as well as characterize our circuit for use in other circuits.
Parts Needed
BCR (CD79A+B (with and without TCS-Gal4VP16), IgM heavy, IgM light), Lyn, Syk-TEVp, (reporter circuit), (whatever is needed to make dox work). We will be testing all combinations of CD79(A/B)-(stop/TCS-Gal4Vp16).
Dox
Setup
All combinations of plasmids being tested will be prepared and put into dox ladders
plasmids v dox>
0
1
10
100
1000
2000
CD79A-stop CD79B-stop
CD79A-tcs-Gal4VP16 CD79B-stop
CD79A-stop CD79B-tcs-Gal4VP1
CD79A-tcs-Gal4VP1 CD79B-tcs-Gal4VP1
First row is control for basal reporter expression.
Expected Results
We should identify what level of Syk-TEVp causes constitutive expression of the reporter and characterize the off behavior of our circuit so we will be able to compare it to the on behavior later.