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This test will vary several circuit parameters to find the ideal sensor and characterize total sensor behavior. |
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This will determine what amounts of Syk-TEVp for each placement of TCS-Gal4VP16 produce the best on/off ratio. |
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BCR (CD79A+B (with and without TCS-Gal4VP16), IgM heavy, IgM light), Lyn, Syk-TEVp, tre-mKate(dox reporter circuit), UAS-eYFP(reporter circuit), hef1a:eBFP(transfection marker), (whatever is needed to make dox work). We will be testing all combinations of CD79(A/B)-(stop/TCS-Gal4Vp16). Dox, beta amyloid (if beta amyloid fails, anti IgM can be used to force the receptor into an active state)
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All combinations of plasmids being tested will be prepared and put into dox ladders
First row is control for basal reporter expression.
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We should identify what level of Syk-TEVp produces the best on/off ratio. Our data will produce a scatter plot of eYFP (reporter) as a function of mKate (driven by the same promoter as syk). |
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