Page History

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TEV cleavage site high or low affinity?

Experiment

-          Western blot HEK293 cells for Syk

• anti-Syk antibody

-          Check BCR membrane localization

• add CD79a/b + mIgM and verify with fluorescent anti-IgM antibodies

-          Check IgM binding

• biotinylated beta-amyloid with fluorescent secondary antibody

-          Check Lyn and Syk membrane localization using fluorescent fusions

• Lyn-YFP
• Syk-mKate

-          Make Syk+TEVp fusions

• Fusions ->

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• Prioritize
  Fusions
  • N terminus: 0 linking aa
  • N terminus: 15 linking aa
  • C terminus: 0 linking aa
  • C terminus: 15 linking aa
  • Inducible expression under TetR -> find upper limit for constitutive behavior

  Syk+TEVp -> find lower limit of beta amyloid (threshold)

   

   

  High Priority Fusions

  -          Syk-BFPeYFP

  -          Lyn-mKATE

  -          CD79A-TCS-Gal4VP16

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