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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 cause excessive constitutive production of reporter so we don't need to test those for the active test. This will also characterize our circuits off behavior so we can compare it to its on behavior later to get the produce the best on/off ratio. There is a test of no Gal4VP16 which will tell us how much constitutive eYFP our promoter produces. |
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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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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 laterproduces 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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